Species Specificity

By using degenerate primers deduced from conserved patterns in the flavivirus polymerase gene, a novel RNA virus was discovered in Rhipicephalus ticks sampled from members of the family Bovidae in Senegal. It was named Ngoye virus (NGOV) after the location from which it was isolated. Viral particles could be observed by electron microscopy, but isolation in vertebrate or invertebrate cell lines or by intracerebral infection of newborn mice remained unsuccessful. This is atypical of recognized arboviruses. The characterization of 4176 nt of the non-structural genes revealed that NGOV is a novel flavivirus species. It forms a distinct phylogenetic lineage related distantly to previously identified members of the genus Flavivirus. Analysis of genetic data suggested that the processing of the NGOV polyprotein and the organization of its replication complex are similar to those of flaviviruses. Together with other recent data, these findings suggest that a large number of viruses related...

The aim of this paper is to desribe Sipunculus (Sipunculus) nudus from the Pakistani marine waters. The knowledge of sipunculans from the Pakistani coast is fragmentary. We can say that Sipuncula is one of the most taxonomically neglected groups of marine invertebrates in Pakistani waters This is the first report of Sipunculus (Sipunculus) nudus collected along the northern Arabian Sea (Pakistan coast).The species at hand is briefly described and photographed The paper additionally comments on its eco morphology, food and medicinal values, distribution, and habitat. A check list of sipunculids already reported from Pakistan is also added.

The histochemical demonstration of nucleoside diphosphatase as a specific microglial marker was used to study the distribution of this glial cell type in the cerebral cortex of Podarcis muralis and Podarcis hispanica. Our results showed that in both species, NDPase staining was specific for the microglial cell population and that microglial cells displayed a specific localization pattern in the different cortical areas. In the medial cortex, microglial cells were principally found in the outer and inner plexiform layers in the strata adjacent to the granular layer. Moreover, some microglial cells were found near the ependymal layer, but no microglial cells were normally present near the brain surface and never in the deep inner plexiform layer. In the dorsomedial cortex, microglial cells were found near the brain surface in the outer plexiform layer, in the upper part of the granular layer, and near the ependymal layer. No microglial cells were found, however, in the outer and inner...

Many important relationships amongst kinetoplastids, including the position of trypanosomatids, remain uncertain, with limited taxon sampling of markers other than small subunit ribosomal RNA (SSUrRNA). We report gene sequences for cytosolic heat shock proteins 90 and/or 70 (HSP90, HSP70) from the potentially early-diverging kinetoplastids Ichthyobodo necator and Rhynchobodo sp., and from bodonid clades '2' (Parabodonidae) and '3' (Eubodonidae). Some of the new cytosolic HSP70 sequences represent a distinct paralog family (HSP70-B), which is related to yet another paralog known from trypanosomatids (HSP70-C). The (HSP70-B, HSP70-C) clade seemingly diverged before the separation between kinetoplastids and diplonemids. Protein phylogenies support the basal placement of Ichthyobodo within kinetoplastids. Unexpectedly, Rhynchobodo usually forms the next most basal group, separated from the clade '1' bodonids with which it has been allied. Bootstrap support is often weak, but the possibility that Rhynchobodo represents a separate early-diverging lineage within core kinetoplastids deserves further testing. Trypanosomatids always fall remote from the root of kinetoplastids, forming a specific relationship with bodonid clades 2 (and 3), generally with strong bootstrap support. These protein trees with improved taxon sampling provide the best evidence to date for a 'late' emergence of trypanosomatids, contradicting recent SSUrRNA-based proposals for a relatively early divergence of this group.

Cystic echinococcosis (CE), caused by Echinococcus granulosus, affects both humans and livestock. Final hosts (dogs) are typically subclinical, while intermediate hosts show symptoms based on cyst location and infection severity. CE is mainly detected in abattoirs via postmortem (PM) examination, making diagnosis challenging and increasing transmission risk. The current study aimed to evaluate the serodiagnosis and statistical significance of using different prepared antigens from cysts against PM method. 170 blood samples were collected from cattle and the separated serum samples were prepared for detection of optical density values by indirect ELISA against different hydatid cysts antigens. The antigens were isolated from various cystic components. Crude hydatid cyst fluid antigen (HCF Ag) was prepared by aspirating fluid from fertile cyst separated from infected organs, centrifuged and aliquoted, laminated layer antigen (LL Ag) obtained by aspirating fluid, separating the laminated layer and homogenized, germinal layer antigen (GL Ag) was isolated through dissection, protoscolices antigen (Pscs Ag) collected, washed with PBS, and all isolates sonicated, and stored at-20°C. Protein concentration was measured using modified Lowry method. The obtained values were statically analyzed. PM prevalence was 11.76%. While ELISA showed 13.5%, with higher prevalence in the liver (7.06%) than lungs (2.35%). Significant differences were noted between antigens, with GL, HCF, and Pscs Ags showing higher values than LL Ag. ELISA diagnostic accuracy was 98.24 % for LL, GL, and Pscs Ags, and 60% for HCF Ag. Our study suggests that GL, HCF, and Pscs Ags were effective and sensitive immunodiagnostic tools for screening hydatosis.

We identified 100 scientific questions that, if answered, would have the greatest impact on conservation practice and policy. Representatives from 21 international organizations, regional sections and working groups of the Society for Conservation Biology, and 12 academics, from all continents except Antarctica, compiled 2291 questions of relevance to conservation of biological diversity worldwide. The questions were gathered from 761 individuals through workshops, email requests, and discussions. Voting by email to short-list questions, followed by a 2-day workshop, was used to derive the final list of 100 questions. Most of the final questions were derived through a process of modification and combination as the workshop progressed. The questions are divided into 12 sections: ecosystem functions and services, climate change, technological change, protected areas, ecosystem management and restoration, terrestrial ecosystems, marine ecosystems, freshwater ecosystems, species management, organizational systems and processes, societal context and change, and impacts of conservation interventions. We anticipate that these questions will help identify new directions for researchers and assist funders in directing funds.

The assembly of fungal communities on stone materials is mainly influenced by the differential bioreceptivity of such materials and environmental conditions. However, little is known about the role of fungal interactions in the colonization and establishment of fungal species. We analyzed the effects of intra- and interspecific interactions between 11 species of fungi in oligotrophic and copiotrophic media and on limestone coupons. In a previous study, these species were the most frequently isolated in the epilithic biofilms of limestone walls exposed to a subtropical climate. In the culture media, we found a greater frequency of intra- and interspecific inhibitory effects in the oligotrophic medium than in the copiotrophic medium. On the limestone coupons, all fungi were able to establish; however, the colonization success rate varied significantly. Cladosporium cladosporioides had a less extensive colonization in isolation (control) than in dual interactions (coexistence) with oth...

The spoT gene of Salmonella typhimurium has been identified. Mutations in spoT map between gltC and pyrE at 79 min. The spoT1 mutant has elevated levels of guanosine 5'-diphosphate-3'-diphosphate (ppGpp) during steady-state growth and exhibits a slower than normal decay of ppGpp after reversal of amino acid starvation. The spoT1 mutation elevates his operon expression but is distinct from known his regulatory mutations. Elevated his operon expression in spoT mutants causes resistance to the histidine analogs, 1,2,4-triazole-3-alanine and 3-amino-1,2,4-triazole. These properties of spoT mutants allowed us to identify and characterize additional spoT mutants. Approximately 40% of these mutants are temperature sensitive for growth on minimal medium, suggesting that the spoT function is essential or that excessive accumulation of ppGpp is lethal.

Fungal spore concentrations vary during day and night. Dry spore types are released during the day, whereas wet spores that have more humidity ratio are released into the air at night. Studies on airborne fungi in Turkey are very rare, and they are daily and weekly monitored only in some cities. The aim of our study was to determine the daily fungal concentrations in Edirne, Turkey. All samples were taken from a level of 11 m. above ground. The sampling time was carried out bihourly in the day, from 8 am to 20 pm, once a week from October 11 to November 15, 2002. Two sampling methods were used for the isolation of fungi: A gravitational and a volumetric method. Statistical analysis was performed in order to determine the relationship between some environmental factors and airborne counts and some correlation were found between the fungal numbers, relative humidity, sampling hours and temperature. A total of 800 fungal colonies in 42 petri dishes were isolated, quantified to determine the frequency of occurrence and then identified as genus level. 9337 cfu/m 3 microfungi were determined by volumetric sampling. Seven fungal genera could be determined, among them Cladosporium and Alternaria were found as the most predominant fungi, followed by Penicillium and Trichoderma.

Stereotypic behavior is an important indicator of compromised welfare. Zoo elephants are documented to perform stereotypic behavior, but the factors that contribute to performance have not been systematically assessed. We collected behavioral data on 89 elephants (47 African [Loxodonta africana], 42 Asian [Elephas maximus]) at 39 North American zoos during the summer and winter. Elephants were videoed for a median of 12 daytime hours per season. A subset of 32 elephants (19 African, 13 Asian) was also observed live for a median of 10.5 nighttime hours. Percentages of visible behavior scans were calculated from five minute instantaneous samples. Stereotypic behavior was the second most commonly performed behavior (after feeding), making up 15.5% of observations during the daytime and 24.8% at nighttime. Negative binomial regression models fitted with generalized estimating equations were used to determine which social, housing, management, life history, and demographic variables were associated with daytime and nighttime stereotypic behavior rates. Species was a significant risk factor in both models, with Asian elephants at greater risk (daytime: p<0.001, Risk Ratio = 4.087; nighttime: p<0.001, Risk Ratio = 8.015). For both species, spending time housed separately (p<0.001, Risk Ratio = 1.009), and having experienced inter-zoo transfers (p<0.001, Risk Ratio = 1.175), increased the risk of performing higher rates of stereotypy during the day, while spending more time with juvenile elephants (p<0.001, Risk Ratio = 0.985), and engaging with zoo staff reduced this risk (p = 0.018, Risk Ratio = 0.988). At night, spending more time in environments with both indoor and outdoor areas (p = 0.013, Risk Ratio = 0.987) and in larger social groups (p = 0.039, Risk Ratio = 0.752) corresponded with reduced risk of performing higher rates of stereotypy, while having experienced inter-zoo transfers (p = 0.033, Risk Ratio = 1.115

Tripartite motif (TRIM) proteins are composed of RING, B-box 2, and coiled coil domains. Some TRIM proteins, such as TRIM5α, also possess a carboxy-terminal B30.2(SPRY) domain and localize to cytoplasmic bodies. TRIM5α has recently been shown to mediate innate intracellular resistance to retroviruses, an activity dependent on the integrity of the B30.2 domain, in particular primate species. An examination of the sequences of several TRIM proteins related to TRIM5 revealed the existence of four variable regions (v1, v2, v3, and v4) in the B30.2 domain. Species-specific variation in TRIM5α was analyzed by amplifying, cloning, and sequencing nonhuman primate TRIM5 orthologs. Lineage-specific expansion and sequential duplication occurred in the TRIM5α B30.2 v1 region in Old World primates and in v3 in New World monkeys. We observed substitution patterns indicative of selection bordering these particular B30.2 domain variable elements. These results suggest that occasional, complex chang...

A high number of tapeworms (Cestoda) of three orders (Caryophyllidea, Bothriocephalidea and Proteocephalidea) have been described from freshwater fish in the Indomalayan Region, with a great majority of them being based on material collected in India (for lists of species,

Seven species of Plakinidae are currently known from Brazil, belonging to genera Plakina (P. trilopha), Plakinastrella (P. microspiculifera, P. onkodes), and Plakortis (P. angulospiculatus, P. halichondrioides, P. insularis, and P. microrhabdifera). In this study we describe four new species of the genera Plakinastrella and Plakortis, increasing the number of plakinid species in Brazil to eleven. Plakortis petrupaulensis sp. nov. is encrusting, light brown; the skeleton is disorganized with few traces of reticulation; lacunae are absent; the spicules are diods and tuberculate microrhabds; spheres are common and triods are absent. Plakortis spinalis sp. nov. is encrusting, gray, cartilaginous, has a reticulated ectosomal skeleton, and is distinguished by the spines in some of the diods and triods; microrhabds are also tuberculate and smooth spheres are common. Plakortis potiguarensis sp. nov. is encrusting, brown with gray tinges, and cartilaginous; the skeleton is disorganized and scarce, composed of thin, irregular diods, triods and relatively large microrhabds. Plakinastrella globularis sp. nov. has a globular shape and dark blue color in vivo; the ectosomal skeleton has a double tangential reticulation and the choanosomal skeleton is reticulated. The spicules are diods, triods, calthrops and microrhabds; spheres are common. Two of the new species have characters previously unknown in their respective genera: the spined diods and triods of Plakortis spinalis sp. nov. and the blue color and microrhabds of Plakinastrella globularis sp. nov. The presence of microrhabds in Plakinastrella suggests a close relationship of this genus with Plakortis.

Ornamental fish constitute a significant sector of the global aquaculture industry, contributing substantial economic value through the aquarium trade, eco-tourism, and export markets. Beyond their aesthetic appeal, they serve as important model organisms in biomedical and developmental research, while also supporting livelihoods in many developing countries. The demand for novel varieties with enhanced colouration, unique morphological traits, and improved resilience to environmental stress has increased markedly in recent years. Conventional selective breeding, although effective, is timeconsuming and limited in its precision. Genome editing technologies, particularly the CRISPR-Cas9 system, have emerged as transformative tools for accelerating trait development in ornamental fishes. CRISPR-Cas9 offers high precision, efficiency, and cost-effectiveness compared to earlier platforms such as transcription activator-like effector nucleases (TALENs) and zinc-finger nucleases (ZFNs). Its application enables targeted modifications of genes influencing pigmentation (e.g., mitfa, slc24a5, kit), fin morphology, growth patterns, and stress tolerance. In addition to generating novel phenotypes of commercial interest, genome editing can improve disease resistance, enhance adaptation to captive environments, and produce sterile strains to mitigate risks of genetic pollution from accidental releases. This review comprises recent advancements in the application of genome editing in ornamental fish.

Crustaceans during a parasitism relationship with fish cause biological disruption and diseases to the hosting fish that threaten their life. Several crustacean groups were reported to parasitize different fish species as definitive hosts. Copepods, Isopods, and Branchiura are the major fish parasitic groups under the crustacea. They are mainly ectoparasites and occasionally, the small and microscopic members can infest the internal organs of their fish hosts. The infected fish are suffering from serious clinical signs and the infested organs are usually suffering from severe pathological disruptions and lesions. Treatment is taking place by several chemical and biological facilities. Here in the current article, the major crustaceans that parasitize on fish are reviewed by describing their corresponding harms to the hosting fish. Factors affecting the prevalence of the crustacean parasites and some of their hazardous effects on the infected fish were considered. In addition, trials for the chemical and biological control of the crustacean infestations are summarized.

Climate change effects on host-parasite interactions have been poorly studied in arid or semiarid habitats. Here, we conducted an experiment aimed to increase the temperature inside European roller Coracias garrulus nest boxes located in a semi-arid habitat on different nest-site types to look for effects on different ectoparasite abundances and nestling growth. Average nest temperature was slightly higher in heated nests than in control nests, although differences were not statistically significant. However, relative humidity was significantly lower at night in heated nests as compared to control nests. The abundance of sand flies, mites and carnid flies was significantly higher in heated, less humid, nests while biting midge abundance was significantly lower in heated nests. Other ectoparasites were not significantly affected by treatment. Relative humidity was high even in heated nests, reaching more than 60%. Sand fly abundance was higher in nests located in sandstone walls, while mite abundance was higher in isolated farmhouses. In addition, sand fly prevalence was higher in nests located in isolated farmhouses and sandstone walls. Heat treatment, nest-site type or ectoparasite abundances did not affect the nestling body mass, wing length or their growth at different nestling ages.

Visceral leishmaniasis (VL) also known as kala-azar is a growing health problem in Ethiopia with an estimated annual VL incidence between 3700 and 7400. The disease is mainly endemic in northwestern parts of the country. The aim of the current study was to determine the sand fly fauna and ecology of Phlebotomus orientalis in two endemic and ecologically distinct areas of northwestern Ethiopia. Sand flies were collected using CDC light traps, sticky traps and pyrethrum spray catches from peri-domestic, mixed forest, farm field and indoor habitats from both Libo-Kemkem (May 2011-April 2012) and Metema (October 2012-September 2013) districts. A total of 51,411 sand fly specimens were collected and identified (10,776 from highland and 40, 635 from the lowland areas). Seven species were found in the highland area: two Phlebotomus spp. (P. orientalis and P. rodhaini) and five Sergentomyia species. Whereas 19 species were found in the lowland area: six Phlebotomus (P. orientalis, P. rodhai...

Snake venoms comprise complex secretions composed of numerous enzymes, peptides, small organic molecules, and inorganic components which, upon injection, induce a broad range of physiological effects determined by their toxin constituents. Snake venom composition varies dramatically among genera and species, and substantial intraspecific variation also occurs in many species. Clinicians report variability in symptomology and in efficacy of antivenom treatment following envenomation by the Northern Pacific and Southern Pacific Rattlesnakes (Crotalus oreganus oreganus and C. o. helleri). This study documents substantial geographic variation in venom protein composition and presence of neurotoxicity among C. oreganus specimens sampled across California, USA. We used multivariate statistics to assess associations of venom composition with phylogenetic distance, geographic distance, and environmental variables (vegetation index, precipitation, and temperature). Environmental factors showed the strongest correlation with venom composition, followed by genetic relatedness, and geographic distance. The findings have direct implications for improved clinical treatment of snakebite, and, potentially, for novel pharmaceutical applications of toxins.

Information on the cetacean fauna found in the nearshore waters of Pagan Island, Commonwealth of the Northern Mariana Islands (CNMI), are reported for the first time. In August 2013 visual and acoustic surveys in the nearshore (< 5.6 kilometers; km [3 nautical miles; nmi]) waters surrounding Pagan Island collected data on cetacean occurrence using line transect (visual and towed acoustic), photo-identification, and moored sonobuoy (passive acoustic monitoring) methods. Three species of cetaceans were visually confirmed at Pagan Island: spinner dolphin (Stenella longirostris), common bottlenose dolphin (Tursiops truncatus), and Cuvier's beaked whale (Ziphius cavirostris). Acoustic encounters included spinner, common bottlenose dolphin, Cuvier's beaked whale, Blainville's beaked whale (Mesoplodon densirostris) and sperm whale (Physeter macrocephalus). The moderately high detection rates of beaked whale species at Pagan can likely be explained by the steep bathymetry and deep-water habitat close to shore. Sperm whales were detected on the acoustic recordings from nighttime moored sonobuoys, indicating their presence within 37 km (20 nmi) of Pagan. On 10 of the 11 research days at Pagan, dolphins were visually or acoustically encountered in the study area, during day and nighttime sampling periods. Photo-identification yielded re-sights of individual spinner dolphins at Pagan Island on successive days. A potential breeding population of bottlenose dolphins was documented at Pagan based on the presence of three calves, including two neonates. The visual and acoustic detection rates for dolphins were relatively low. Caution should be used in interpreting any of the detection rate results due to the small sample sizes. The density or abundance of cetaceans in the study area could not be estimated because of small sample sizes. If future surveys are conducted using similar methods the line transect survey data could be pooled to estimate density and abundance.

Three distinct bamboo bat species (Tylonycteris) are known to inhabit tropical and subtropical areas of Asia, i.e., T. pachypus, T. robustula, and T. pygmaeus. This study performed karyotypic examinations of 4 specimens from southern Chinese T. p. fulvidus populations and one specimen from Thai T. p. fulvidus population, which detected distinct karyotypes (2n=30) compared with previous karyotypic descriptions of T. p. pachypus (2n=46) and T. robustula (2n=32) from Malaysia. This finding suggested a cryptic Tylonycteris species within T. pachypus complex in China and Thailand. Morphometric studies indicated the difficulty in distinguishing the cryptic species and T. p. pachypus from Indonesia apart from the external measurements, which might be the reason for their historical misidentification. Based on 623 bp mtDNA COI segments, a phylogeographic examination including T. pachypus individuals from China and nearby regions, i.e., Vietnam, Laos, and Cambodia, was conducted to examine t...

Species-specific urokinase receptor (uPAR) ligands with improved pharmacokinetics were generated by sitespecific mutagenesis and amino-terminal pegylation. These molecules were used to probe the role of uPAR in brain tumor progression and angiogenesis. The ligands blocked endothelial cell tube formation in Matrigel in a species-specific manner and reduced both baseline and uPA amino-terminal fragment-stimulated cell migration on vitronectin gradients. Treatment of U87MG gliomas implanted orthotopically in mice with single speciesspecific or combination uPAR ligands resulted in significant decreases in tumor size, which translated to increases in survival time, and which were most significant when the murine-specific ligand was included. Further analysis of tumors showed that the reduced sizes were correlated with a decrease in tumor cell proliferation and mean vessel density and an increase in tumor cell apoptosis. In addition, a large increase in collagen deposition was observed in the treated groups. Statistical analysis showed that the combination therapy demonstrated a clear synergy as compared to the individual agent treatments. These results suggest that the major role of the uPAR system in brain tumor progression is in the stromal compartment and particularly in neovascularization, a hallmark of invasive brain tumors.

The complex geological and climatic history of the Neotropics has had major implications on the diversification of plant lineages. Chrysobalanaceae is a pantropical family of trees and shrubs with 75% of its 531 species found in the Neotropics, and a time-calibrated phylogeny of this family should shed light on the tempo of diversification in the Neotropical flora. Previously published phylogenetic hypotheses of this family were poorly supported, and its biogeography remains unclear. We assembled the complete plastid genome of 51 Chrysobalanaceae species, and increased taxon sampling by Sanger-sequencing of five plastid regions for an additional 88 species. We generated a time-calibrated tree including all 139 Chrsyobalanaceae species and 23 outgroups. We then conducted an ancestral area reconstruction analysis and estimated diversification rates in the family. The tree generated with the plastid genome alignment was almost fully resolved. It supports the polyphyly of Licania and Hi...

SUMMARYWe quantitatively assessed ocular vascular patterns of six Antarctic notothenioid fishes that vary in their expression of the circulating oxygen-binding protein, hemoglobin (Hb). Digital image analyses revealed marked differences in vessel morphometries among notothenioid species. Hemoglobinless (–Hb) icefishes display mean vessel length densities that are greater (Chaenocephalus aceratus, 5.51±0.32 mm mm–2; Champsocephalus gunnari, 5.15±0.50 mm mm–2) than those observed in red-blooded (+Hb) species(Gymnodraco acuticeps, 5.20±0.46 mm mm–2; Parachaenichthyes charcoti, 4.40±0.30 mm mm–2; Trematomus hansoni, 3.94±0.08 mm mm–2; Notothenia coriiceps, 2.48±0.21 mm mm–2). –Hb fishes also have mean vessel diameters that are ∼1.5 times greater than vessel diameters of +Hb species (–Hb,0.193±0.006 mm; +Hb, 0.125±0.005 mm). Vascular density index(VDI), a stereological index that is affected by both vessel number and length, is greatest in –Hb C. aceratus (3.51±0.20) and lowest in +Hb N....

Eelpouts of the family Zoarcidae comprise a monophyletic group of marine fishes with a worldwide distribution. Centers of high zoarcid diversity occur in the North Atlantic and North Pacific, with important radiations into the Arctic, along southern South America, and into the Southern Ocean around Antarctica. Along with snailfishes (Liparidae), zoarcids form an important component of the non-notothenioid fauna in the subzero shelf waters of Antarctica. We document the anatomy and histology of the brains, cranial nerves, olfactory apparatus, cephalic lateral lines, taste buds, and retinas of three Antarctic zoarcid species, living at depths of 310 -939 m, representing three of the nine genera from this region. The primary emphasis is on Ophthalmolycus amberensis, and we provide a detailed drawing of the brain and cranial nerves of this species. Although this brain reflects general perciform neural morphology, it exhibits a reduction of the (optic) tecta and the eminentia granulares and crista cerebellares of the lateral line system. Interspecific differences among the three species are slight. The olfactory rosette consists of three to four lamellae and the nasal sac, contrary to the claim of Fanta et al. ([2001] Antarct Rec, Natl Inst Polar Res, Tokyo 45:27-42), is not in communication with the cephalic lateral line system. Primary olfactory neurons are abundant and converge on branches of the olfactory nerve. Numerous taste buds are located in the lips. All three species lack an ocular choroid rete and have relatively thin retinas with a low cell density and a single bank of rods as the only type of photoreceptor. Neural diversification among Antarctic zoarcids has not involved the evolution of sensory specialists; brain and sensory organ morphologies do not approach the condition seen in primary deep-sea fishes, or even that of some sympatric non-perciform secondary deep-sea fishes, including liparids and muraenolepidids (eel cods). There may be phylogenetic constraints on brain morphology in perciforms such that we do not see extreme specialization in sensory and neural systems for deep habitats. We suggest that the brains and sensory organs of Antarctic zoarcids reflect habitation of 500 -2,000-m depths and likely reflect morphologies seen in zoarcids living on continental slopes elsewhere in the world. This balance among the sensory modalities makes zoarcids relatively generalized among secondary deep-sea fishes and may be one of the reasons this opportunistic and adaptable group has been successful in colonizing a variety of emergent and ephemeral habitats.

This study investigates the effect of biofortification of Granny Smith apple (Malus domestica) agro-industrial byproducts (RM-N) through combined abiotic stresses (UVA radiation and wounding) to enhance their phenolic compound content and bioactivities. The applied abiotic stress conditions significantly increased the total phenolic content and the content of procyanidin B2, catechin, and epicatechin, with a maximum increase of 305 % in biofortified tissue (RM-E) compared to untreated samples (RM-N). Simulated gastrointestinal digestion improved the bioaccessibility of key phenolic compounds in RM-E, enhancing their antioxidant capacity by 22.8 % and the inhibition of nitric oxide (NO) production by 107 % in lipopolysaccharide (LPS)-stimulated murine macrophages (RAW 264.7). Furthermore, RM-E displayed selective antiproliferative activity against colorectal cancer cell lines (Caco-2, HT29), achieving lower IC 50 values than RM-N samples, while maintaining low cytotoxicity in healthy fibroblast cells. These findings demonstrate the feasibility of using combined abiotic stresses as a sustainable and cost-effective strategy to valorise apple by-products, providing bioactive-rich ingredients with potential applications in functional foods and nutraceuticals.

Used as markers of postnatal growth closure sequences of 22 ectocranial sutures and synchondroses were recorded in a sample of 1161 skulls belonging to 38 species from all major placental clades: Afrotheria, Xenarthra, Laurasiatheria and Euarchontoglires (Boreoeutheria). The maximum closure level, which is not significantly correlated to body mass, is higher in Afrotheria and Xenarthra than in Boreoeutheria. Only the basioccipito-basisphenoid and the basioccipito-exoccipital synchondroses close in all species sampled, the supraoccipito-exoccipital and the inter-parietal sutures do in most species. Parsimov retrieved more heterochronic shifts for Afrotheria and Xenarthra than for Boreoeutheria. The amount of intraspecific variation differs among the species sampled being high among xenarthran species and low among afrotherians. Specimens (162) representing 12 marsupial genera were also analysed. Placentals exhibit a larger number of suture closures than marsupials and in both groups the sutures at the base of the skull are the first to fuse starting with the basioccipito-exoccipital.

At present, despite extensive laboratory investigations, most cases of porcine abortion remain without an etiological diagnosis. Due to a lack of recent data on the abortigenic effect of order Chlamydiales, 286 fetuses and their placentae of 113 abortion cases (1-5 fetuses per abortion case) were investigated by polymerase chain reaction (PCR) methods for family Chlamydiaceae and selected Chlamydia-like organisms such as Parachlamydia acanthamoebae and Waddlia chondrophila. In 0.35% of the cases (1/286 fetuses), the Chlamydiaceae real-time PCR was positive. In the Chlamydiaceae-positive fetus, Chlamydia abortus was detected by a commercial microarray and 16S ribosomal RNA PCR followed by sequencing. The positive fetus had a Porcine circovirus-2 coinfection. By the Parachlamydia real-time PCR, 3.5% (10/286 fetuses of 9 abortion cases) were questionable positive (threshold cycle values: 35.0-45.0). In 2 of these 10 cases, a confirmation by Chlamydiales-specific real-time PCR was possible. All samples tested negative by the Waddlia real-time PCR. It seems unlikely that Chlamydiaceae, Parachlamydia, and Waddlia play an important role as abortigenic agents in Swiss sows.

This document provides detailed findings on the reproductive biology, behaviour, and ecological adaptations of the Indian chameleon (Chamaeleo zeylanicus) based on studies conducted at Satkoshia Gorge Sanctuary (1975-1981). Key points include: 1. Reproductive Biology • Mating: Occurs in late August, preceded by courtship involving "chase and escape" behaviour. Males display territorial aggression, while females initially resist but become receptive briefly.

Objective: To investigate species-specific envenoming rates and spectrum of severity of funnel-web spider bites, and the efficacy and adverse effects of funnel-web spider antivenom. Data sources: Cases were identified from a prospective study of spider bite presenting to four major hospitals and three state poisons information centres (1999-2003); museum records of spider specimens since 1926; NSW Poisons Information Centre database; MEDLINE and EMBASE search; clinical toxinology textbooks; the media; and the manufacturer's reports of antivenom use. Data extraction: Patient age and sex, geographical location, month, expert identification of the spider, clinical effects and management; envenoming was classified as severe, mildmoderate or minor/local effects. Data synthesis: 198 potential funnel-web spider bites were identified: 138 were definite (spider expertly identified to species or genus), and 77 produced severe envenoming. All species-identified severe cases were attributed to one of six species restricted to NSW and southern Queensland. Rates of severe envenoming were: Hadronyche cerberea (75%), H. formidabilis (63%), Atrax robustus (17%), Hadronyche sp. 14 (17%), H. infensa (14%) and H. versuta (11%). Antivenom was used in 75 patients, including 22 children (median dose, 3 ampoules; range, 1-17), with a complete response in 97% of expertly identified cases. Three adverse reactions were reported, all in adults: two early allergic reactions (one mild and one with severe systemic effects requiring adrenaline), and one case of serum sickness. Conclusions: Severe funnel-web spider envenoming is confined to NSW and southern Queensland; tree-dwelling funnel webs (H. cerberea and H. formidabilis) have the highest envenoming rates. Funnel-web spider antivenom appears MJA 2005; 182: 407-411 effective and safe; severe allergic reactions are uncommon.

Eighteen species belonging to the Carex genus were checked for the presence and the amount of eight phenolic acids (p-hydroxybenzoic, vanillic, caffeic, syringic, protocatechuic, p-coumaric, sinapic, and ferulic) by means of HPLC. Both the free and bonded phenolic acids were analyzed. The majority of the analyzed acids occurred in the studied species in relatively high amounts. The highest concentrations found were caffeic acid and p-coumaric acid, for which the detected levels were negatively correlated. A very interesting feature was the occurrence of sinapic acid, a compound very rarely detected in plant tissues. Its distribution across the analyzed set of species can be hypothetically connected with the humidity of plants&#39; habitats. Several attempted tests of aggregative cluster analysis showed no similarity to the real taxonomical structure of the genus Carex. Thus, the phenolic acids&#39; composition cannot be considered as the major taxonomical feature for the genus Carex.

The DNA sequence of the intragenic region of the rat 45S ribosomal RNA precursor was determined. This sequence contains 2282 nucleotides and extends from the conserved EcoR I site near the 3' terminus of 18S rRNA to 69 nucleotides downstream of the 5' terminus of 28S rRNA. The sequences corresponding to 18S and 5.8S rRNA were identified by comparison with previously published data. The 5' terminus of rat 28S rRNA was identified by SI nuclease protection and reverse transcriptase elongation assays. The internal transcribed spacers were found to be 1066 and 765 nucleotides long and had little homology with those of Xenopus and yeast. Regions of sequence homology between rat and Xenopus were found at the junctions of the internal transcribed spacers with 18S, 5.8S and 28S rRNA. These homologies suggest that these sequences may function as recognition sites for the processing of the ribosomal precursor RNA.

xa s .ed u , m rya n @ m a il .ute xas .edu 2 Te xas M em o ria l M useum , University o f Te xa s , Austin , Te xas 78705 , US A ; E -m a il : ca t® sh @ m a il .ute xas .edu Ab stra ct .Ð C h a ra cters de rive d fr om adve rtise m e n t calls, m o rph ology, allozym e s, an d the se q ue n ce s of th e sm a ll su b u n it of the m itoch o n d rial rib osom al ge n e (12S ) an d th e cytoch rom e o xida se I (C O I) m itoch o n drial ge n e we re use d to e stim a te the ph yloge n y o f fro gs of th e Ph ysala em us p ustulo sus grou p (Le ptod actylidae ). T h e co m b ina b ility o f th e se d ata pa rtition s wa s a sse sse d in se veral ways : m e asure s of ph yloge n e tic sign al, ch ara cter su pp ort fo r tre e s, congru e n c e of tre e topologie s, co m patib ility o f da ta p artitions with sub optim a l tree s, a n d h om oge n e ity of da ta partition s. C o m b ine d parsim on y an a lysis o f a ll data e q ua lly we igh ted yielde d th e sam e tre e as th e 12S pa rtition an alyzed un de r pa rsim on y a n d m axim u m like lih ood . T h e C O I, allozym e , an d m orph ology partition s we re ge n e rally c on gru e n t an d co m patib le with the tree de rived fro m co m b ine d d ata . T h e call d ata we re sign i® can tly diƒe re n t fro m all oth e r p artitions, wh e the r co n side re d in te rm s o f tree top ology alon e , p artition h o m oge n e ity, o r co mp atib ility o f d ata with tre e s de rive d fro m o the r p artitions. Th e la ck of e ƒect o f th e call d ata on th e top ology of the c om b in e d tre e is prob ab ly d ue to th e sm all n u m b e r of ca ll ch aracte rs. T h e ge n e ra l inco n grue n ce o f the call d ata with othe r d ata p artition s is con siste n t with th e ide a tha t th e adve rtise m e n t c alls of th is grou p o f fr og s are u n d e r stron g se xual se le ction. [ Ad vertise m e n t calls ; b e h a vior ; com b ine d-da ta a n alysis ; d ata partition s ; fro gs ; L e ptoda ctylidae ; Ph y sa la em us ; se n sory e xp loitation h yp oth e sis.] W he the r or n ot to com b in e da ta sets has be e n discusse d wid ely in the re cen t litera ture (

A simple molecular tool was developed and tested to identify seven mealybug species found in North American vineyards: Pseudococcus maritimus Ehrhorn, Pseudococcus viburni (Signoret), Pseudococcus longispinus (Targioni-Tozzeti), Pseudococcus calceolariae (Maskell), Planococcus ficus (Signoret), Planococcus citri (Risso), and Ferrisia gilli Gullan. The developed multiplex PCR is based on the mitochondrial cytochrome c oxidase subunit one gene. In tests, this single-step multiplex PCR correctly identiÞed 95 of 95 mealybug samples, representing all seven species and collected from diverse geographic regions. To test the sensitivity, single specimen samples with different Pl. ficus developmental stages (egg to adult female and adult male) were processed PCR and the resulting output provided consistent positive identiÞcation. To test the utility of this protocol for adult males caught in sex baited pheromone traps, Pl. ficus adult males were placed in pheromone traps, aged at a constant temperature of 26 Ϯ 2ЊC, and processed with the multiplex each day thereafter for 8 d. Results showed consistent positive identiÞcation for up to 6 d (range, 6 Ð 8 d). Results are discussed with respect to the usefulness of this molecular tool for the identiÞcation of mealybugs in pest management programs and biosecurity of invasive mealybugs.

Les levures du genre Dekkera/Brettanomyces sont reconnues comme les agents de contamination des vins rouges responsables des défauts phénolés et animaux. Une procédure complète de dépistage et d'isolement a été développée en associant plusieurs critères discriminants. Elle a permis de confirmer la présence du contaminant tout au long du procédé, de la matière première (raisins sur pieds), au produit fini (vin en bouteilles). Des études de terrain, en amont du procédé de vinification ont mis en avant l'existence de profils hétérogènes de présence sur les parcelles de vigne du fait de paramètres environnementaux. Des facteurs à la vigne, tels que l'humidité, influent sur la présence du contaminant sur les baies de raisin. Des études en laboratoire sur des souches isolées de Brettanomyces bruxellensis, ont révélé une très grande diversité au sein de l'espèce. Cette variabilité intra-spécifique s'exprime à différents niveaux : génétique, phénotypique et physiologiques. Les profils de croissance, la consommation des substrats (carboné et azoté), la production de 4-éthyl-phénol et d'acide acétique ont constitué autant d'éléments de différenciation entre les souches étudiées. Nous avons par ailleurs montré que l'emploi d'adjuvants nologiques n'est pas sans conséquence sur le développement de la levure et son activité contaminante. Ainsi la thiamine (à raison de 6 mg/hL), l'azote ajouté sous forme de sulfate d'ammonium, ou encore les tanins nologiques (de raisin et de bois) favorisent dans une certaine mesure le développement de Brettanomyces. Pour chacun de ces adjuvants les effets sur la croissance et la production d'éthyl-phénol sont différents.

Les levures du genre Dekkera/Brettanomyces sont reconnues comme les agents de contamination des vins rouges responsables des défauts phénolés et animaux. Une procédure complète de dépistage et d'isolement a été développée en associant plusieurs critères discriminants. Elle a permis de confirmer la présence du contaminant tout au long du procédé, de la matière première (raisins sur pieds), au produit fini (vin en bouteilles). Des études de terrain, en amont du procédé de vinification ont mis en avant l'existence de profils hétérogènes de présence sur les parcelles de vigne du fait de paramètres environnementaux. Des facteurs à la vigne, tels que l'humidité, influent sur la présence du contaminant sur les baies de raisin. Des études en laboratoire sur des souches isolées de Brettanomyces bruxellensis, ont révélé une très grande diversité au sein de l'espèce. Cette variabilité intra-spécifique s'exprime à différents niveaux : génétique, phénotypique et physiologiques. Les profils de croissance, la consommation des substrats (carboné et azoté), la production de 4-éthyl-phénol et d'acide acétique ont constitué autant d'éléments de différenciation entre les souches étudiées. Nous avons par ailleurs montré que l'emploi d'adjuvants nologiques n'est pas sans conséquence sur le développement de la levure et son activité contaminante. Ainsi la thiamine (à raison de 6 mg/hL), l'azote ajouté sous forme de sulfate d'ammonium, ou encore les tanins nologiques (de raisin et de bois) favorisent dans une certaine mesure le développement de Brettanomyces. Pour chacun de ces adjuvants les effets sur la croissance et la production d'éthyl-phénol sont différents.

Schistosomes are digenean flukes, parasitic of birds, mammals and crocodiles. The family Schistosomatidae contains species of considerable medical and veterinary importance, which cause the disease schistosomiasis. Previous studies, both morphological and molecular, which have provided a good deal of information on the phylogenetics of this group, have been limited in the number of species investigated or the type or extent of molecular data used. This paper presents the most comprehensive phylogeny to date, based on the sequences of 3 genes, complete ribosomal small subunit rRNA and large ribosomal subunit rRNA, and mitochondrial cytochrome oxidase 1, sequenced from 30 taxa including at least 1 representative from 10 of the 13 known genera of the Schistosomatidae and 17 of the 20 recognized Schistosoma species. The phylogeny is examined using morphological characters, intermediate and definitive host associations and biogeography. Theories as to the origins and spread of Schistosom...

Sand dune quarries are a location of common aggregate mining activity developed in coastal areas, especially in the southeast Buenos Aires province, Argentina. In this article, spontaneous plant development after extraction activity ceased was evaluated. Five areas (three quarried and two natural/conservation areas) were sampled for plant cover and composition as well as sediment characterization. Different indexes, principal component analysis, and cluster analyses were applied to compare the areas. The dominant families observed in four of the five areas were Asteraceae, Poaceae, and Cyperaceae, and most of the species are commonly found in sandy and humid soils and/or modified/anthropized ones. Percentages of plant cover increased with time because of the cessation of active aggregate extraction. Indexes and multivariate analyses showed that it was possible to distinguish quarried and natural areas based on composition and vegetation cover. The distribution of plant species among the four areas responded to the presence of mining activity, but it also responded to the topographical position and consequently the depth of the groundwater level. Besides these differences, the four areas shared many native species. The results might indicate that once the activity has ceased, quarried areas may spontaneously and quickly develop a plant community with some similarities to those present in the nonquarried areas. However, given that the extracting activity involves the removal of the soil, revegetation of this type of environment depends on the presence of natural areas in the surroundings, which can serve as a source of seeds and propagules for plant regeneration.

SummaryPrimula sieboldii E. Morren is a perennial clonal herb that is widely distributed in Japan, but in danger of extinction in the wild. In a previous study, we revealed the genetic diversity of the species using chloroplast and nuclear DNA and used this information to define conservation units. However, we lacked information on adaptive genetic diversity, which is important for long-term survival and, thus, for the definition of conservation units. In order to identify adaptive traits that showed adaptive differentiation among populations, we studied the genetic variation in six quantitative traits within and among populations for 3 years in a common garden using 110 genets from five natural populations from three regions of Japan. The number of days to bud initiation was adaptive quantitative trait for which the degree of genetic differentiation among populations (QST) was considerably larger than that in eight microsatellite markers (FST). The relationship between this trait a...

The ceramide (Cer) and sphingomyelin (SM) species of cultured differentiated rat cerebellar granule cells and human fibroblasts were characterized by electrospray ionization-mass spectrometry. We identified 35 different species of Cer and 18 species of SM in human fibroblasts, and 35 different species of Cer and 9 species of SM were characterized in rat neurons. The main Cer species of rat cerebellar granule cells contained d18:1 sphingosine linked with palmitic, stearic, or nervonic fatty acid, and the two main SM species were d18:1,16:0 and d18:1,18:0. Both sphingolipids were enriched in detergent-resistant membranes (DRMs; or lipid rafts), and significant differences were found in the sphingolipid patterns of DRMs and of detergentsoluble fractions (DSF) from these cells. In human fibro-

Scanning electron microscopy (SEM) has been used as a tool to facilitate the morphological description of mites and to complement the information obtained from the light microscope. Unfortunately, several of SEM techniques used for soft-bodied mites such as Tyrophagus, have in many cases not shown the desired results. This investigation sought to develop an efficient procedure for preparing specimens of T. putrescentiae for SEM so that its major morphological characters could be observed and photographed. This species is the most important contaminant of tissue culture laboratories in Costa Rica. Seven methods were used to process the samples. The evaluated treatments showed differences in their ability to preserve the morphological structures of the specimens. The advantages and disadvantages of each treatment are discussed. Mites processed using ethanol were the most viable option with method 3 being the procedure that showed the best results; this technique also reduced the time and costs of processing the specimens.

The white matter of the brain consists of fiber tracts that connect different regions of the brain. Among these tracts, the intrahemispheric cortico-cortical connections are called association fibers. The U-fibers are short association fibers that connect adjacent gyri. These fibers were thought to work as part of the cortico-cortical networks to execute associative brain functions. However, their anatomy and functions have not been documented in detail for the human brain. In past studies, U-fibers have been characterized in the human brain with diffusion tensor imaging (DTI). However, the validity of such findings remains unclear. In this study, DTI of the macaque brain was performed, and the anatomy of U-fibers was compared with that of the human brain reported in a previous study. The macaque brain was chosen because it is the most commonly used animal model for exploring cognitive functions and the U-fibers of the macaque brain have been already identified by axonal tracing studies, which makes it an ideal system for confirming the DTI findings. Ten U-fibers found in the macaque brain were also identified in the human brain, with a similar organization and topology. The delineation of these species-conserved white matter structures may provide new options for understanding brain anatomy and function.

1. In degraded tropical landscapes, lack of seed dispersal can strongly limit recovery, and restoration interventions can overcome this barrier by attracting dispersers. However, seed dispersal patterns are typically studied over short time periods, thus the influences of temporal and spatial variability on seed arrival cannot be teased apart.
2. The choice of management approach can have important implications for restoration-mediated community reassembly. Accordingly, we used a 3.5-year record of seed deposition in pre-montane tropical wet forest in southern Costa Rica to examine how seed arrival differed between passive (natural regeneration) and active (applied nucleation, plantation) restoration after a decade of recovery, compared to remnant forest. We investigated: (a) how restoration treatments affected seed deposition rates and community composition; (b) how within-plot heterogeneity of animal-dispersed seed deposition varied by intervention; and (c) how interannual variation influenced animal-dispersed seed arrival across treatments.
3. Overall seed rain composition and diversity in restoration treatments was converging towards, but still differed substantially from, remnant forest (89.7%, 86.6% and 76.3% Shannon diversity recovered in applied nucleation, plantation and natural regeneration respectively).
4. Within-plot animal-dispersed seed heterogeneity was similar in applied nucleation and remnant forest, 27.0% more heterogeneous in applied nucleation than plantation, and equivalent when comparing natural regeneration to either applied nucleation or plantation.
5. In contrast to active interventions, animal-dispersed tree and shrub communities did not differ year to year in natural regeneration, which may promote the assembly of relatively homogeneous plant communities at this successional stage.
6. Synthesis and applications. Compared to natural regeneration, active restoration interventions: (a) catalysed the recovery of seed diversity (overall Shannon diversity 17.5% and 13.4% higher in applied nucleation and plantation respectively), (b) shifted seed community composition towards remnant forest more rapidly (overall Shannon diversity 13.4% and 10.2% closer), (c) almost doubled the proportion of later-successional tree species arriving, and (d) had seed communities that differed year to year—a pattern not observed in natural regeneration. Finally, applied nucleation was the only intervention where seed arrival was as spatially heterogeneous as remnant forest, highlighting that this approach may facilitate the recovery of specific natural dispersal processes.

Questions: Vascular epiphytes constitute a large proportion of tropical forest plant biodiversity, but are among the slowest plants to recolonize secondary forests. We asked whether tree planting for ecological restoration accelerates epiphyte community recovery. Does the spatial configuration of tree planting matter? What landscape contexts are most suitable for epiphyte restoration?

Location: Restored pastures in premontane Coto Brus County, Puntarenas,
Costa Rica.

Methods: We surveyed vascular epiphyte species growing on the lower
trunks of 1083 trees in 13 experimental restoration sites. Each site contained three 0.25-ha treatment plots: natural regeneration, trees planted in patches or ‘islands’ and tree plantations. Sites spanned elevational (1100–1430 m) and deforestation (4–94% forest cover within a 100-m radius around each site) gradients.

Results: Vascular epiphytes were twice as diverse in planted restoration plots (islands and plantations) as in natural regeneration; we observed this at the scale of individual host trees and within 0.25-ha treatment plots. Contributing factors included that trees in planted restoration plots were larger, older, more abundant and composed of different species than trees in naturally regenerating plots. Epiphyte species richness increased with surrounding forest cover within 100– 150 m of restoration plots. Epiphyte communities were also twice as diverse at higher (1330–1430 m) vs lower (1100–1290 m) elevation sites. Epiphyte groups responded differently to restoration treatments and landscape factors; ferns were responsible for higher species richness in planted restoration plots, whereas angiosperms drove elevation and forest cover effects.

Conclusions: Tree planting for ecological restoration enriched epiphyte communities compared to natural regeneration, likely because planted forests contained more, bigger and older trees. Tree island plantings were equally effective compared to larger and more expensive plantations. Restoration sites nearer to existing forests had richer epiphyte recolonization, likely because nearby forests provisioned restoration sites with angiosperm seeds. Collectively, results suggest that restoration practitioners can enrich epiphyte community development by planting trees in areas with higher surrounding forest cover, particularly at higher elevations.

Information on the parasites of salmonids in Japanese waters that was published during the years 1889-1986 is assembled in the form of Parasite-Host and Host-Parasite lists with accompanying bibliography. Ninety-four named species of parasites (18 Protozoa, 5

A comprehensive inventory of global biodiversity would be greatly improved by automating methods for species delimitation. The Automatic Barcode Gap Discovery method, the Poisson tree processes algorithm and the Generalized mixed Yule-coalescent model have been proposed as means of increasing the rate of biodiversity description using single locus data. We applied these methods to explore the diversity within the Aglaopheniidae, a hydrozoan family with many species widely distributed across tropical and temperate oceans. Our analyses revealed widespread cryptic diversity in this family, almost half of the morpho-species presenting several independent evolutionary lineages, as well as support for cases of synonymy. For two common species of this family, Lytocarpia brevirostris and Macrorhynchia phoenicea, we compared the outputs to clustering analyses based on microsatellite data and to nuclear gene phylogenies. For L. brevirostris, microsatellite data were congruent with results of ...

Complement resistance is an important virulence trait of Yersinia enterocolitica (Ye). The predominant virulence factor expressed by Ye is Yersinia adhesin A (YadA), which enables bacterial attachment to host cells and extracellular matrix and additionally allows the acquisition of soluble serum factors. The serum glycoprotein vitronectin (Vn) acts as an inhibitory regulator of the terminal complement complex by inhibiting the lytic pore formation. Here, we show YadA-mediated direct interaction of Ye with Vn and investigated the role of this Vn binding during mouse infection in vivo. Using different Yersinia strains, we identified a short stretch in the YadA head domain of Ye O:9 E40, similar to the &#39;uptake region&#39; of Y. pseudotuberculosis YPIII YadA, as crucial for efficient Vn binding. Using recombinant fragments of Vn, we found the C-terminal part of Vn, including heparin-binding domain 3, to be responsible for binding to YadA. Moreover, we found that Vn bound to the bact...

The temperature-sensitive BHK21 hamster cell line tsBN67 ceases to proliferate at the nonpermissive temperature after a lag of one to a few cell divisions, and the arrested cells display a gene expression pattern similar to that of serum-starved cells. The temperature-sensitive phenotype is reversible and results from a single missense mutation-proline to serine at position 134-in HCF, a cellular protein that, together with the viral protein VP16, activates transcription of herpes simplex virus (HSV) immediate-early genes. The tsBN67 HCF mutation also prevents VP16 activation of transcription at the nonpermissive temperature. The finding that the same point mutation in HCF disrupts both VP16 function and the cell cycle suggests that HCF plays a role in cell-cycle progression in addition to VP16-dependent transcription.