Species- and sex-specific chemical composition from an internal gland-like tissue of an African frog family.

Intraspecific chemical communication in frogs is understudied and the few published cases are limited to externally visible and male-specific breeding glands. Frogs of the family Odontobatrachidae, a West African endemic complex of five morphologically cryptic species, have large, fatty gland-like strands along their lower mandible. We investigated the general anatomy of this gland-like strand and analysed its chemical composition. We found the strand to be present in males and females of all species. The strand varies in markedness, with well-developed strands usually found in reproductively active individuals. The strands are situated under particularly thin skin sections, the vocal sac in male frogs and a respective area in females. Gas-chromatography/mass spectrometry and multivariate analysis revealed that the strands contain sex- and species-specific chemical profiles, which are consistent across geographically distant populations. The profiles varied between reproductive and non-reproductive individuals. These results indicate that the mandibular strands in the Odontobatrachidae comprise a so far overlooked structure (potentially a gland) that most likely plays a role in the mating and/or breeding behaviour of the five Odontobatrachus species. Our results highlight the relevance of multimodal signalling in anurans, and indicate that chemical communication in frogs may not be restricted to sexually dimorphic, apparent skin glands.

Variants in FtsJ RNA 2'-O-Methyltransferase 3 and Growth Hormone 1 are associated with small body size and a dental anomaly in dogs.

Domesticated dogs show unparalleled diversity in body size across breeds, but within breeds variation is limited by selective breeding. Many heritable diseases of dogs are found among breeds of similar sizes, suggesting that as in humans, alleles governing growth have pleiotropic effects. Here, we conducted independent genome-wide association studies in the small Shetland Sheepdog breed and discovered a locus on chromosome 9 that is associated with a dental abnormality called maxillary canine-tooth mesioversion (MCM) (P = 1.53 × 10-7) as well as two body size traits: height (P = 1.67 × 10-5) and weight (P = 1.16 × 10-7). Using whole-genome resequencing data, we identified variants in two proximal genes: FTSJ3, encoding an RNA methyltransferase, and GH1, encoding growth hormone. A substitution in FTSJ3 and a splice donor insertion in GH1 are strongly associated with MCM and reduced body size in Shetland Sheepdogs. We demonstrated in vitro that the GH1 variant leads to exon 3 skipping, predicting a mutant protein known to cause human pituitary dwarfism. Statistical modeling, however, indicates that the FTSJ3 variant is the stronger predictor of MCM and that each derived allele reduces body size by about 1 inch and 5 pounds. In a survey of 224 breeds, both FTSJ3 and GH1 variants are frequent among very small "toy" breeds and absent from larger breeds. Our findings indicate that a chromosome 9 locus harboring tightly linked variants in FTSJ3 and GH1 reduces growth in the Shetland Sheepdog and toy breed dogs and confers risk for MCM through vertical pleiotropy.

Comparative analysis of swine leukocyte antigen gene diversity in European farmed pigs.

In Europe, swine represent economically important farm animals and furthermore have become a preferred preclinical large animal model for biomedical studies, transplantation and regenerative medicine research. The need for typing of the swine leukocyte antigen (SLA) is increasing with the expanded use of pigs as models for human diseases and organ-transplantation experiments and their use in infection studies and for design of veterinary vaccines. In this study, we characterised the SLA class I (SLA-1, SLA-2, SLA-3) and class II (DRB1, DQB1, DQA) genes of 549 farmed pigs representing nine commercial pig lines by low-resolution (Lr) SLA haplotyping. In total, 50 class I and 37 class II haplotypes were identified in the studied cohort. The most common SLA class I haplotypes Lr-04.0 (SLA-1*04XX-SLA-3*04XX(04:04)-SLA-2*04XX) and Lr-32.0 (SLA-1*07XX-SLA-3*04XX(04:04)-SLA-2*02XX) occurred at frequencies of 11.02 and 8.20% respectively. For SLA class II, the most prevalent haplotypes Lr-0.15b (DRB1*04XX(04:05/04:06)-DQB1*02XX(02:02)-DQA*02XX) and Lr-0.12 (DRB1*06XX-DQB1*07XX-DQA*01XX) occurred at frequencies of 14.37 and 12.46% respectively. Meanwhile, our laboratory has contributed to several vaccine correlation studies (e.g. Porcine Reproductive and Respiratory Syndrome Virus, Classical Swine Fever Virus, Foot-and-Mouth Disease Virus and Swine Influenza A Virus) elucidating the immunodominance in the T-cell response with antigen specificity dependent on certain SLA-I and SLA-II haplotypes. Moreover, these SLA-immune response correlations could facilitate tailored vaccine development, as SLA-I Lr-04.0 and Lr-32.0 as well as SLA-II Lr-0.15b and Lr-0.12 are highly abundant haplotypes in European farmed pigs.

Immune gene expression and genome-wide association analysis in rainbow trout with different resistance to Yersinia ruckeri infection.

Selective breeding programmes involving marker assisted selection of innately pathogen resistant strains of rainbow trout rely on reliable controlled infection studies, extensive DNA typing of individual fish and recording of expression of relevant genes. We exposed juvenile rainbow trout (6 h bath to 2.6 × 105 CFU mL-1) to the fish pathogen Yersinia ruckeri serotype O1, biotype 2, eliciting Enteric Red Mouth Disease ERM, and followed the disease progression over 21 days. Cumulative mortality reached 42% at 12 days post challenge (dpc) after which no disease signs were recorded. All fish were sampled for DNA-typing (50 k SNP chip, Affymetrix®) throughout the course of infection when they showed clinical signs of disease (susceptible fish) or at day 21 when fish showed no clinical signs of disease (survivors - resistant fish). Genome-wide association analyses of 1027 trout applying single nucleotide polymorphisms (SNPs) as markers revealed an association between traits (susceptible/resistant) and certain regions of the trout genome. It was indicated that multiple genes are involved in rainbow trout resistance towards ERM whereby it is considered a polygenic trait. A corresponding trout group was kept as non-exposed controls and a comparative expression analysis of central innate and adaptive immune genes in gills, spleen and liver was performed for three fish groups: 1) moribund trout exhibiting clinical signs 7 dpc (CS), 2) exposed fish without clinical signs at the same sampling point (NCS) and 3) surviving fish at 21 dpc (survivors). Immune genes encoding inflammatory cytokines (IL-1ß, IL-2A, IL-6A, IL-8, IL-10A, IL-12, IL-17A/F2A, IL-17C1, IL-17C2, IL-22, IFNγ, TNFα), acute phase reactants (SAA, C3, cathelicidins, lysozyme) were expressed differently in CS and NCS fish. Correlation (negative or positive) between expression of genes and bacterial load suggested involvement of immune genes in protection. Down-regulation of adaptive immune genes including IgDm, IgDs, IgT and TCR-ß was seen primarily in CS and NCS fish whereas survivors showed up-regulation of effector molecule genes such as cathelicidins, complement and lysozyme suggesting their role in clearing the infection. In conclusion, SNP analyses indicated that ERM resistance in rainbow trout is a multi-locus trait. The gene expression in surviving fish suggested that several immune genes are associated with the trait conferring resistance.

Isolation and Characterization of Feeding-Deficient Strains in Inbred Lines of the Hydrozoan Jellyfish Cladonema pacificum.

Feeding behavior in cnidarians has been studied as a model experimental system in physiology and neurobiology. Although the feeding response in cnidarians, such as Hydra, is triggered by chemical signals, the underlying molecular mechanisms that ensure their precise execution are not well understood. It could be largely due to the lack of genetic analysis in cnidarian experimental systems. Cladonema pacificum is a hydrozoan jellyfish that is easy to maintain and cross for genetic analysis in the laboratory. To establish C. pacificum as a model experimental animal in cnidarians, we have been inbreeding strains of jellyfish. Here, we document our progress in developing C. pacificum inbred lines and feeding-defective strains that we isolated in the course of inbreeding. In the inbred lines, an increasing number of feeding-defective strains appeared as descending generations and finally all the F5 progeny showed a feeding-deficient phenotype presumably owing to inbreeding depression. Feeding behaviors of these strains were analyzed by video microscopy and we found that the feeding-defective strains captured prey, but could not kill them. After trapping prey, wild-type medusae contracted their tentacles tightly and then bent the tentacles to bring the prey to the mouth; however, feeding-defective medusae rarely contracted their tentacles and did not bend. These feeding-defective phenotypes are caused by lack of stinging nematocytes in their tentacle batteries. These findings furnish a clue to the regulatory aspects of feeding behavior, but also reveal the mechanisms of stinging nematocyte transport in tentacles.

A spatial model for the beneficial use of dredge spoil deposition: Creation and management of breeding habitat for reddish egrets in Texas.

Many species of coastal waterbirds have faced population declines in recent years as a result of loss and degradation of habitat. The creation of new habitat through strategic placement of dredge spoil can be an effective tool to mitigate habitat loss. The use of dredge spoil islands for nesting, wintering, and migratory stopover habitat by waterbirds has been documented extensively over the past several decades. Most of the reddish egret (Egretta rufescens) breeding population in the United States occurs in Texas, where it has declined by more than 30% since the 1970s. Reddish egrets breed on dredge spoil islands along the Texas coast, and the stability of breeding colonies has been variable since the 1970s. If nesting habitat is degraded or limiting in Texas, the creation of new nesting islands through strategic dredge spoil placement could benefit reddish egrets. We developed a spatially explicit prioritization model for targeted conservation of reddish egrets in the Laguna Madre, Texas, through colonial island establishment using dredge spoil. Prioritization is based on availability of foraging areas, potential competition at foraging areas, and distance of potential sites to the mainland. 'Priority areas' are potential nesting island locations that would maximize the foraging benefits for breeding reddish egrets. Model outputs show that highest priority areas in the Laguna Madre are located in the southernmost portion of the lagoon. Other priority areas are located in the lower Laguna Madre, and at the mouth of Baffin Bay. In addition, we provide a repeatable and transparent framework for the development of spatial support tools that help guide targeted placement of dredge spoil in an effort to conserve colonial waterbirds. We illustrate the model development for reddish egrets, a threatened species in Texas, however our framework can be applied to other similar species.

Morphological diversity of integumentary traits in fowl domestication: Insights from disparity analysis and embryonic development.

The domestication of the fowl resulted in a large diversity of integumental structures in chicken breeds. Several integumental traits have been investigated from a developmental genetics perspective. However, their distribution among breeds and their developmental morphology remain unexplored. We constructed a discrete trait-breed matrix and conducted a disparity analysis to investigate the variation of these structures in chicken breeds; 20 integumental traits of 72 chicken breeds and the red junglefowl were assessed. The analyses resulted in slight groupings of breed types comparable to standard breed classification based on artificial selection and chicken type use. The red junglefowl groups together with bantams and European breeds. We provide new data on the red junglefowl and four chicken breeds, demonstrating where and when variation arises during embryonic development. We document variation in developmental timing of the egg tooth and feather formation, as well as other kinds of developmental patterning as in the anlagen of different type of combs. Changes in epithelial-mesenchymal signaling interactions may drive the highly diverse integument in chickens. Experimental and comparative work has revealed that the cranial neural crest mesenchyme mediates its interactions with the overlying epithelium and is the likely source of patterning that generates diversity in integumental structures.

Breeder-reported patterns of antimicrobial use and point prevalence of methicillin-resistant Staphylococcus spp. among breeding bitches in the southwestern United States.

Methicillin-resistant Staphylococcus pseudintermedius skin infections are an increasing concern in veterinary medicine, especially when found in juvenile dogs with no prior antimicrobial exposure. The objectives of this study were to assess the prevalence of methicillin-resistant staphylococci in breeding bitches and survey antimicrobial administration by their breeders. A total of 17 breeders and 54 bitches were included. Bitches were housed in diverse environments throughout the Phoenix, Arizona, USA metropolitan region. Nasal and oral gingival swabs were submitted for selective culture. Methicillin-resistant Staphylococcus aureus was not present, while methicillin-resistant Staphylococcus pseudintermedius was found in only 1/54 (1.9%) dogs. Survey results revealed that 16 bitches, which did not include the 1 MRSP-positive dog, had received antimicrobials within 6 months prior to sampling. Regardless of the low point prevalence identified, veterinarians and breeders should be cognizant of risks for bacterial resistance with the overuse of antimicrobials.

Patron d'utilisation des antimicrobiens rapporté par les éleveurs et prévalence ponctuelle de Staphylococcus spp. résistant à la méthicilline parmi des chiennes de reproduction dans le Sud-Ouest des États-Unis. Les infections cutanées dues à Staphylococcus pseudintermedius résistant à la méthicilline sont une préoccupation grandissante en médecine vétérinaire, surtout lorsque retrouvée chez des chiens juvéniles sans exposition préalable aux antibiotiques. Les objectifs de la présente étude étaient d'évaluer la prévalence de staphylocoques résistants à la méthicilline chez des chiennes reproductrices et sonder l'administration d'antimicrobiens par les éleveurs. Un total de 17 éleveurs et de 54 chiennes furent inclus. Les chiennes étaient logées dans divers environnements de la région métropolitaine de Phoenix, Arizona, USA. Des écouvillons du nez et des gencives orales furent soumis pour une culture sélective. Aucun Staphylococcus aureus résistant à la méthicilline ne fut détecté, alors que S. pseudintermedius fut trouvé chez seulement 1/54 chiens (1,9 %). Les résultats du sondage ont révélé que 16 chiennes, qui n'incluaient pas la chienne positive pour MRSP, avaient reçu des antimicrobiens à l'intérieur des 6 mois précédant l'échantillonnage. Indépendamment de la faible prévalence ponctuelle identifiée, les vétérinaires et les éleveurs devraient être au fait des risques pour la résistance bactérienne associée avec la surutilisation des antimicrobiens.(Traduit par Dr Serge Messier).

The Saliva Proteome of Dogs: Variations Within and Between Breeds and Between Species.

Saliva is a complex multifunctional fluid that bathes the oral cavity to assist in soft and hard tissue maintenance, lubrication, buffering, defense against microbes, and initiating digestion of foods. It has been extensively characterized in humans but its protein composition in dogs remains poorly characterized, yet saliva composition could explain (patho) physiological differences between individuals, breeds and with humans. This pilot discovery study aimed to characterize canine saliva from two breeds, Labrador retrievers and Beagles, and to compare this with human saliva using quantitative mass spectrometry. The analysis demonstrated considerable inter-individual variation and difference between breeds; however these were small in comparison to the differences between species. Functional mapping suggested roles of detected proteins similar to those found in human saliva with the exception of the initiation of digestion as salivary amylase was lacking or at very low abundance in canine saliva samples. Many potential anti-microbial proteins were detected agreeing with the notion that the oral cavity is under continuous microbial challenge.

A frameshift variant in the EDA gene in Dachshunds with X-linked hypohidrotic ectodermal dysplasia.

X-linked hypohidrotic ectodermal dysplasia (XLHED) is a genetic disease characterized by hypoplasia or absence of hair, teeth and sweat glands. The EDA gene, located on the X chromosome, encodes the type II transmembrane protein ectodysplasin A. Variants in the EDA gene can lead to XLHED in humans, mice, cattle and dogs. In the present study, we investigated a litter of Dachshund puppies, of which four male puppies showed clinical signs of XLHED. We performed a candidate gene analysis in one affected puppy and several non-affected relatives. This analysis revealed a single base-pair deletion in the coding sequence of the EDA gene in the affected puppy (NM_001014770.2:c.842delT). The deletion is predicted to cause a frameshift, NP_001014770.1:p.(Leu281HisfsTer22), leading to a premature stop codon which truncates more than one quarter of the EDA protein. Sanger sequencing results confirmed that this variant was inherited from the dam. Based on knowledge about the functional impact of EDA variants in dogs and other species, c.842delT is a convincing candidate causative variant for the observed XLHED in the male puppies.

Apical Elongation of Molar Teeth in Captive Microtus Voles.

Molar apical elongation (MAE) was the leading cause for euthanasia or death in a captive breeding colony of endangered Amargosa voles ( Microtus californicus scirpensis). Clinical signs included ocular discharge, abnormal mastication, dyspnea, abnormal mentation, weight loss, and death. Although the severity varied, all molars in all quadrants were affected. When severe, the overgrown molar reserve crown and apex protruded into the nasal meatuses, the orbit, the calvarial vault and through the ventral margin of the mandible. Overall prevalence in the colony was 63% (92/146 voles) and increased to 77% in aged voles (>1 year). Mean age of onset was 5.3 months (1.7-11.2 months). Progression to extreme severity occurred over 1 to 3 months. Mean survival was 10.9 months (7.1-21.7 months). Histologically, the lesion was characterized by odontogenic hyperplasia and dysplasia. MAE was also documented in museum specimens of 2 other M. californicus subspecies ( M. californicus californicus, M. californicus vallicola) and 3 other Microtus species ( M. montanus, M. pennsylvanicus, M. socialis). In the M. californicus californicus collection, overall prevalence was 35.1% (129/368 skulls) and increased to 77.3% in aged voles (>1 year). A probable genetic influence was identified in the museum collection of M. californicus californicus. The etiopathogenesis of MAE is likely multifactorial, due to (1) inherent continuous odontogenic proliferation, (2) inadequate occlusal attrition, and (3) possible heritable disease susceptibility. In captivity, dietary or other management of occlusal attrition to prevent or delay MAE is a fundamental concern.

Associations between liveweight, body condition score and previous reproductive outcomes, and the risk of ewes bred at 18-months of age being dry at docking.

AIMS Firstly, to investigate associations between liveweight and body condition score (BCS) of two-tooth ewes (18-months-old at breeding) at breeding, pregnancy diagnosis (PD) and pre-lambing and the risk of being dry at docking, on commercial New Zealand sheep farms. Secondly, to investigate the association between previous reproductive outcomes as ewe lambs, and risk of being dry at docking as two-tooth ewes. METHODS Two-tooth ewes (n=9,006) were enrolled in four cohorts from three commercial sheep farms between 2010-14. Ewes were weighed and BCS assessed immediately pre-breeding, at PD (mid-pregnancy) and pre-lambing. At PD, ewes were identified as either non-pregnant, or having single or multiple fetuses. Palpation and examination of udders at docking was used to classify each ewe as either lactating or dry at docking. RESULTS Overall, 437/8,025 (5.4%) of ewes that were diagnosed pregnant at PD were dry at docking. The risk of being dry at docking decreased with increasing pre-lambing conceptus adjusted liveweight (CALW) on all farms (p≤0.002); for 2010-born ewes from Farm A the OR=0.87 (95% CI=0.81-0.92); for Farm B the OR=0.88 (95% CI=0.83-0.92) and for Farm C the OR=0.86 (95% CI=0.79-0.95). The risk of being dry at docking also decreased with increasing CALW gain from PD to pre-lambing for all farms (p≤0.003); for 2010-born ewes from Farm A the OR=0.89 (95% CI=0.84-0.94); for Farm B the OR=0.85 (95% CI=0.81-0.89) and for Farm C the OR=0.88 (95% CI=0.80-0.96). There was no association between BCS at breeding, PD or pre-lambing and the risk of being dry at docking for 2010-born ewes from Farm A, Farm B or Farm C (p>0.05). For 2010-born ewes on Farm A, the risk of being dry at docking was greater for two-tooth ewes that were previously dry at docking as ewe lambs than those that were lactating at docking as ewe lambs (OR=1.7 (95% CI=1.1-2.8); p=0.018), but this difference was not observed for ewes on Farm B or Farm C (p>0.5). CONCLUSIONS There were negative associations between ewe CALW pre-lambing, and CALW gain between PD and pre-lambing, and risk of being dry at docking. For all cohorts, heavier ewes and those that gained CALW were less likely to be dry at docking than lighter ewes or those that lost CALW, however these relationships varied between cohorts.

An investigation on the impact of substrate type, temperature, and iodine on moon jellyfish production.

Jellyfish are a popular public aquarium species, however, their collection from natural populations is undesirable due to impact on species abundance and bycatch. Thus, a sustainable supply of jellyfish bred in-house would be highly desirable. Here we describe an investigation into developing a sustainable moon jellyfish, Aurelia aurita, breeding program by determining the impact of substrate type on reattachment of polyps and the influence of iodine and temperature on strobilation and ephyra production. To test whether reattachment and growth of moon jellyfish polyps are influenced by substrate type polyps were provided with anthropogenic and natural substrates after being dislodged in experimental aquaria. Polyps selectively re-attached to plastics rather than natural materials. However, polyp growth was similar on all tested substrates. We tested whether cooling and addition of iodine affected strobilation. A period of cooling of around 10 °C while also introducing soluble iodine to the polyps enhanced strobilation rate. This method produced ephyra at a reliable rate in captivity negating the need for collection of wild individuals providing a supply of individuals for exhibit and for conservation research within a public aquarium. These results demonstrate that plastics should be adopted as an easier to colonize substrate and the use of cooling with iodine addition can enhance sustainable breeding protocols of moon jellyfish and may be relevant to the production of comparable jellyfish species.

Breeding experiments and genome-wide association analysis elucidate two genetically different forms of non-syndromic congenital cleft lip and jaw in Vorderwald × Montbéliarde cattle.

Non-syndromic congenital cleft lip and jaw (CLJ) is a condition reported in Vorderwald × Montbéliarde cattle. The objective of the present study was to perform a genome-wide association study (GWAS) for 10 CLJ-affected and 50 unaffected Vorderwald × Montbéliarde cattle using the bovine Illumina high density bead chip to identify loci for this condition. Phenotypic classification of CLJ was based on a detailed recording of orofacial structures using computed tomography. A breeding experiment among CLJ-affected Vorderwald × Montbéliarde cattle and CLJ-affected Vorderwald × Montbéliarde cattle with unaffected Holsteins confirmed recessive inheritance and different loci for bilateral or left-sided versus right-sided CLJ. The GWAS for the five cases with right-sided CLJ gave a genome-wide signal on bovine chromosome (BTA) 29 at 16 Mb. For the four left-sided and one bilateral CLJ case, a genome-wide significant association was identified on BTA4 at 32 Mb. Two different loci are very likely to be involved in CLJ in Vorderwald × Montbéliarde cattle because experimental matings among affected cows and bulls with different types of CLJ did not result in CLJ-affected progeny, and in addition, two different loci were also found through GWAS and mapped on two different bovine chromosomes. Validation in 346 Vorderwald × Montbéliarde cattle for the highly associated SNPs on BTA4 and 29 gave ratios of 33/346 (0.095, BTA4) and 6/346 (0.017, BTA29) homozygous mutant genotypes. Further studies should elucidate the responsible mutations underlying the different types of CLJ in Vorderwald × Montbéliarde cattle.

Transcriptome analysis reveals the genetic basis underlying the seasonal development of keratinized nuptial spines in Leptobrachium boringii.

BACKGROUND: The expression of sexually selected traits often varies with populations' breeding cycles in many animals. The elucidation of mechanisms underlying the expression of such traits is a research topic in evolutionary biology; however, the genetic basis of the seasonal development of their expression remains unknown. Male Leptobrachium boringii develop keratinized nuptial spines on their upper jaw during the breeding season that fall off when the breeding season ends. To illuminate the genetic basis for the expression of this trait and its seasonal development, we assessed the de novo transcriptome for L. boringii using brain, testis and upper jaw skin and compared gene expression profiles of these tissues between two critical periods of the spine growth cycle. RESULTS: We identified 94,900 unigenes in our transcriptome. Among them, 2,131 genes were differentially expressed between the breeding period when the spines developed and the post-breeding period when the spines were sloughed. An increased number of differentially expressed genes (DEGs) were identified in the upper jaw skin compared with the testis and brain. In the upper jaw skin, DEGs were mainly enriched in cytosolic part, peptidase inhibitor activity and peptidase regulator activity based on GO enrichment analysis and in glycolysis/gluconeogenesis, ribosome biogenesis in eukaryotes and retinol metabolism based on KEGG enrichment analysis. In the other two tissues, DEGs were primarily involved in the cell cycle, DNA replication and melatonin production. Specifically, insulin/insulin-like growth factor and sex steroid hormone-related DEGs were identified in the upper jaw skin, indicating . The expression variation of IGF2 and estrogen-related genes may be the main factors regulating the seasonal development of the spines. CONCLUSIONS: Our study provides a list of potential genes involved in the regulation of seasonal development of nuptial spines in L. boringii. This is the first transcriptome survey of seasonally developed sexually selected traits for non-model amphibian species, and candidate genes provided here may provide valuable information for further studies of L. boringii.

Influence of sperm fertilising concentration, sperm selection method and sperm capacitation procedure on the incidence of numerical chromosomal abnormalities in IVF early bovine embryos.

The occurrence of numerical chromosomal aberrations, widely described as a major cause of mortality in in vitro-produced (IVP) embryos, has been linked to several factors. In the present study we investigated the effect of sperm fertilising concentration and semen handling (sperm selection and capacitation) before IVF on the rate of numerical chromosomal abnormalities in bovine embryos. In all, 466 IVP cattle embryos were karyotyped throughout three sequential experiments, analysing the effects of sperm fertilising concentration (0.1, 1.0 or 10×10(6) spermatozoa mL(-1)), selection method (unselected or Percoll-selected spermatozoa) and capacitation medium (bovine serum albumin (BSA), heparin or their combination). The percentage of normal (diploid) and aberrant (haploid, polyploid or aneuploid) embryos was noted in each experiment. The rate of numerical chromosomal abnormalities was mainly affected by sperm fertilising concentration (P<0.01) and, to a lesser extent, by the sperm capacitation medium (P<0.05). Polyploidy and haploidy rates were only affected by sperm fertilising concentration (P<0.05). Interestingly, the sperm selection technique used in the present study did not reduce the incidence of chromosome abnormalities in IVP cattle embryos (P>0.05). Finally, aneuploidy rates were not affected during the experiments (P>0.05), which suggests that they are not related to sperm-related factors. On the basis of these results, we conclude that sperm fertilising concentration is the 'paternal' key factor that affects the rate of numerical chromosomal abnormalities in IVP bovine embryos. By making small adjustments to fertilising protocols, the rate of cytogenetically aberrant embryos can be markedly reduced.

Marker-assisted selection in breeding silkworm strains with high tolerance to fluoride, scaleless wings, and high silk production.

In the silkworm (Bombyx mori), tolerance to fluoride and scaleless wings are controlled by the dominant gene Dtf (dominant tolerance to fluoride) and recessive gene nlw (no Lepidoptera wings), respectively, and these genes have been mapped by using simple sequence repeat and sequence tag site markers. Marker-assisted evaluation and selection of silkworms with fluoride tolerance and scaleless wings were used for predicting fluoride resistance and scaleless wings in backcrossed animals. A silkworm strain was bred using this method, and its economic characteristics were found to be similar to those of commercial silkworms. These methods will therefore be useful for silkworm breeding programs and in screening for two or more characteristics of interest for segregating populations.

The use of noninvasive and minimally invasive methods in endocrinology for threatened mammalian species conservation.

Endocrinology is an indispensable tool in threatened species research. The study of endocrinology in threatened species not only advances knowledge of endocrine mechanism but also contributes to conservation efforts of studied species. To this end, endocrinology has been traditionally used to understand reproductive and adrenocortical endocrine axes by quantifying excreted steroid metabolites. From these studies a large body of knowledge was created that contributed to the field of endocrinology, aided conservation efforts, and created a template by which to validate and conduct this research for other species. In this regard noninvasive hormone monitoring has become a favored approach to study the basic endocrinology of wildlife species. Due to the increased understanding of endocrine physiology of threatened species, breeding rates of captive population have improved to levels allowing for reintroduction of species to restored natural ecosystems. Although these approaches are still employed, advances in biochemical, molecular, and genomic technologies are providing inroads to describe lesser known endocrine activity in threatened species. These new avenues of research will allow for growth of the field with greater depth and breadth. However, for all approaches to endocrinology, limitations on resources and access to animals will require innovation of current methodologies to permit broad application for use in threatened species research.

Medically important mosquitoes in the rubber plantation belt of central Kerala, India.

Entomological surveys were carried out in the rubber plantation belt of Kerala to record mosquito fauna. Samples were collected from 23 randomly selected localities using standard methods for a period of three years, from Feb- ruary 2008 to January 2011. Thirty-two species belonging to nine genera: Aedes, Anopheles, Armigeres, Coquillettidia, Culex, Heizmannia, Mansonia, Toxorhynchites, and Uranotaenia were recorded. Many of the recorded species were medically im- portant as potential vectors of dengue fever, chikungunya, Japanese encephalitis, malaria and filariasis.

Geometric morphometric evaluation of mandibles of four sheep breeds: Bardoka, Ivesi, Polish Mountain sheep and Turcana.

The enduring relationship between humans and domestic sheep has evolved over millennia, showcasing diverse uses such as meat, milk, wool, leather and fur, shaped by geographical, historical, cultural and social factors. The sheep breeds discussed include the Ivesi from Southeastern Anatolia, known for its varied animal products; the resilient Turcana breed of Romania; Kosovo's Bardoka, valued for its triple-purpose characteristics; and Poland's Polish Mountain Sheep, uniquely utilized for milk production in cheese making. Sheep, with their enduring relationship with humans and significant economic importance, have attracted scientific interest in morphometric studies of their mandibles, yielding valuable data applicable across various fields including basic anatomy, veterinary clinical anatomy, zooarchaeology and veterinary forensic medicine. Traditional morphometric studies rely on statistical methods to compare length, depth and angular ratios between anatomical formations, often highlighting differences between specific points but not fully revealing shape variations between distinct groups. Geometric morphometric analysis has emerged as a preferred method in recent years, enabling shape analyses using coordinate data from various imaging techniques, facilitating a comprehensive examination of mandibular morphometrics among sheep breeds across different countries. This study involved four sheep breeds from different countries, namely Ivesi from Turkey, Bardoka from Kosovo, Polish Mountain Sheep from Poland and Turcana from Romania, with a total of 70 mandibles sourced from various veterinary faculties. Mandibular photographs were meticulously captured, focusing on the right side of mandible pairs and placing landmarks and semi-landmarks along the entire edge, enabling geometric morphometric analysis using tpsUtil, tpsDig2 and MorphoJ software. The analysis included principal component analysis, canonical variate analysis and discriminant function analysis for pairwise comparisons, facilitating a comprehensive examination of mandibular shape variations among the different sheep breeds. Using geometric morphometric methods, this study analysed mandibles from four distinct sheep breeds sourced from different countries, revealing notable variations in regions such as the ramus mandibula, angulus mandibula and incisive areas, attributed to genetic, geographical and dietary influences, highlighting the importance of continued research to better comprehend these shape differences.