Genomic regions and enrichment analyses associated with carcass composition indicator traits in Nellore cattle

The aim of this study was to estimate genetic parameters and identify genomic regions associated with carcass traits obtained by ultrasound and visual scores in Nellore cattle. Data from ~66,000 animals from the National Association of Breeders and Researchers (ANCP) were used. The variance components for backfat thickness, rump fat thickness and Longissimus muscle area (LMA) were estimated considering a linear model whereas a threshold model for body structure (BS), finishing precocity (FP) and musculature (MS) traits. The SNP solutions were estimated using the ssGBLUP approach by considering windows of 10 consecutive SNPs. Regions that accounted for more than 1.0% of the additive genetic variance were used. Genes identified within the significant windows, such as FOXA3, AP2S1, FKRP, NPASI and ATP6V1G1, were found to be related with MS, while OMA1 and FFGY with BS and FP traits. The PLTP, TNNC2 and GPAT2 genes were found in the regions associated with LMA, as well as TKT, FNDC5 and CHRND can strongly be related with fat deposition. Gene enrichment analysis revealed processes that might be directly influenced the organism growth and development. These results should help to better understand the genetic and physiological mechanisms regulating growth and body composition, muscle tissue development and subcutaneous fat expression, and this information might be useful for future genomic studies in Nellore cattle.     

Mycobacterium avium subsp. paratuberculosis in cow bulk tank milk in Cyprus detected by culture and quantitative IS900 and F57 real-time PCR

The possibility that Mycobacterium avium subsp. paratuberculosis (MAP) plays some role in the development of Crohn's disease in humans is attracting attention to milk and milk products originating from infected animals. In this study, we focused on the detection of MAP in 220 bulk tank milk (BTM) samples from all dairy cattle herds in Cyprus. In total, 63 (28.6%) BTM milk samples were found to be positive for MAP using quantitative real-time PCR assays for IS900 and F57. The presence of MAP in BTM was low, and was assessed to be several tens of MAP cells per one ml of BTM. Milk samples examined by cultivation were found to be negative for MAP in all 220 BTM. In two BTM samples cultivation and subsequent sequencing of 16S rRNA revealed two isolates of M. fortuitum.     

Determination of Phylogenetic Relationships of Flavobacterium psychrophilum (Flexibacter psychrophilus), Flavobacterium columnare (Flexibacter columnaris), and Flexibacter maritimus by Sequence Analysis of 16S Ribosomal RNA Genes Amplified by Polymerase Chain Reaction

Abstract

The nearly complete small subunit 16S ribosomal RNA (rRNA) gene sequence amplified by polymerase chain reaction was determined for Flavobacterium psychrophilum (formerly Flexibacter psychrophilus) by using automated nucleotide sequencing. The sequence was found to be 1,465 base pairs (bp) in length, a size consistent with previously determined sequences for 14 other bacterial species from various taxa, including the yellow-pigmented bacteria. Sequence signatures confirmed that this organism was a member of the bacterial division Bacteroides–Flavobacterium. Parsimonious and additive phylogenetic trees were constructed with homology, and pairwise evolutionary distances were used to estimate phylogenetic relationships. Data show that F. psychrophilum, F. columnare, and Flexibacter maritimus are closely related, have a common descent, and represent a distinct group within the division Bacteroides–Flavobacterium. This group also included other organisms from the genera Flavobacterium and Cytophaga. Further, Flavobacterium aquatile, the type species for the genus Flavobacterium, was also determined to be a member of this “Flavobacterium–Cytophaga–Flexibacter subcomplex.” This supports previous assertions that the type strain of Flavobacterium should be changed to a more representative species such as Flavobacterium breve.     

DNA barcoding for identification of cryptic species in the field and existing museum collections: a case study of Aethomys and Micaelamys (Rodentia: Muridae)

DNA barcoding has been proposed as a method for species identification. However, this method has been criticised for its over-reliance on a single mitochondrial gene. In this study, four mitochondrial gene regions and one nuclear gene region were used to investigate their different abilities to identify tissue associated with museum specimens of Aethomys chrysophilus, Aethomys ineptus and Micaelamys namaquensis. Aethomys chrysophilus and the more recently elevated A. ineptus are indistinguishable on morphological grounds; however, their ranges are largely parapatric with only one syntopic locality currently known. All of the mitochondrial gene regions were able to separate M. namaquensis from A. chrysophilus and A. ineptus, but they varied in their abilities to resolve differences between A. chrysophilus and A. ineptus. The sequence results identified a specimen from KwaZulu-Natal that was misclassified and should have been identified as A. ineptus. Seven specimens that had not been reclassified following the elevation of A. ineptus to species level were identified as A. ineptus. Individuals of A. chrysophilus from Malawi could not be classified as either A. chrysophilus or A. ineptus, and may be a hybrid or a new, distinct species. This study indicates that DNA barcoding may be used to separate M. namaquensis from A. chrysophilus and A. ineptus, and although it was not able to separate A. chrysophilus and A. ineptus, it did indicate specimens from Malawi may be a new cryptic species.     

Presence of Ureaplasma diversum in the Australian cattle population

In cattle, Ureaplasma diversum has been associated with decreased fertility, granular vulvovaginitis, endometritis, salpingitis and spontaneous abortion in cows and seminal vesiculitis, balanoposthitis and changes in bull sperm. The presence of U. diversum within the Australian cattle population has not been established. One of the aims of this study was to determine if U. diversum was present in Australian cattle, using culture and polymerase chain reaction (PCR), both of which are considered to be gold standards for bacterial identification. Of 100 samples collected from 66 male and 34 female cattle, 15 were positive for U. diversum. Therefore, Australia can no longer be considered free of U. diversum. Further studies should be conducted to ascertain the effects of U. diversum within Australian cattle herds and, if warranted, to investigate prevention, treatment and eradication protocols.     

The rough-toothed dolphin genome provides new insights into the genetic mechanism of its rough teeth

The rough-toothed dolphin (Steno bredanensis) is characterized by having teeth covered in finely wrinkled vertical ridges, which is a general manifestation of amelogenesis imperfecta. The rough surfaces are hypothesized to be an evolutionary morphological trait of feeding adaptation to increase the dolphin's grip on prey. Here, we assembled a rough-toothed dolphin genome and performed the comparative genomic analysis to reveal the genetic basis of the special enamel. Results showed that genes related to enamel development or dental diseases have undergone diversified adaptive changes that may shape the special enamel morphology of this dolphin species, including positive selection (CLDN19, PRKCE, SSUH2, and WDR72), rapid evolution (LAMB3), or unique amino acid substitutions (AMTN, ENAM, MMP20, and KLK4). Meanwhile, the historical demography of rough-toothed dolphin indicated several distinct population fluctuations associated with climate change. The genome-wide heterozygosity of this dolphin is in the middle of all published data for cetaceans. Although the population is considerable, there may be population or subspecies differentiation, and with the global warming and the increasing disturbance of human activities, we should pay more attention to protection in the future. Together, our study brings new insights into the genetic mechanisms that may have driven the evolution of the special enamel morphology in rough-toothed dolphins and provides the first results of genetic heterozygosity and population historical dynamics of this species, which have important guiding implications for the conservation of this dolphin species.     

Detection of Coxiella burnetii in buffaloes aborted fetuses by IS111 DNA amplification: A preliminary report

The aim of this study was to evaluate by PCR analyses the presence of Coxiella burnetii infection in fetuses of water buffalo (Bubalus bubalis) in the Campania region (Southern Italy). Samples were collected only from aborted fetuses and C. burnetii presence was evaluated by one-tube nested PCR amplification of the IS111 repetitive element. Of the 164 fetuses examined 14 (17.5%) were positive after DNA amplification, showing that C. burnetii occurs in this population of water buffaloes.However, more extensive prevalence studies need to be carried out to define the role of buffaloes as reservoirs for this pathogen and also the role of C. burnetii as an abortive agent in this animal.     

A serologic study on Toxoplasma gondii infection in slaughtered sheep and goats in Qazvin Province,Iran

Background

Toxoplasma gondii is a common protozoan parasite among all mammals, in particular small ruminants, worldwide. Traditional husbandry can be a major risk factor for infection of sheep and goats with this parasite.

Objectives

The present study aimed to determine the current status of the prevalence for T. gondii in livestock of Qazvin Province.

Methods

In this cross-sectional study, the sera of 455 sheep and 375 goats were examined to detect anti-Toxoplasma IgG antibodies by using in-house indirect ELISA.

Results

Overall, 33.62% (153/455) of sheep and 36.41% (130/375) of goats were positive for anti-Toxoplasma IgG antibodies with no statistically significant difference. The prevalence rate of T. gondii among the sheep of Qazvin County was significantly higher than in Abyek and Abhar counties (p < 0.001).

Conclusions

The results of the present study indicate that the prevalence of T. gondii in sheep and goats of the study area is high. Therefore, the meat of the animals reared in this area can be a potential source of human infections by this parasite.

     

Identification of Rickettsia felis in fleas but not ticks on stray cats and dogs and the evidence of Rickettsia rhipicephali only in adult stage of Rhipicephalus sanguineus and Rhipicephalus haemaphysaloides

Rickettsia spp. are zoonotic pathogens and mainly transmitted by various arthropod vectors, such as fleas, ticks, and lice. Previous epidemiological studies indicated that ectoparasites infested on dogs or cats may be infected by Rickettsia spp., and transmit them to human beings accidentally. In this study, the prevalence of Rickettsia infection was evaluated using fleas and ticks from stray dogs and cats in Taiwan. A total of 158 pools made by 451 cat fleas (Ctenocephalides felis) from 37 dogs and 4 cats were used for analysis. Besides, 386 Rhipicephalus ticks collected from the other 62 stray dogs were included in this study. Nymphal and adult ticks were individually analyzed but larvae were separated into 21 pools for molecular detection. Partial sequencing analysis of the gltA gene was applied for Rickettsia identification. The results showed that 44.3% (70/158) of the cat flea pools were harboring Rickettsia DNA. Although 6.9% (13/187) of adult ticks were infected with Rickettsia, neither larval pools nor nymphal ticks were found to contain Rickettsia DNA. According to the results of sequencing analyses, all Rickettsia PCR-positive cat flea pools were infected with R. felis, and all Rickettsia PCR-positive adult ticks were infected with R. rhipicephali. The results of this study demonstrated that C. felis but not Rhipicephlus sanguineus (the brown dog tick) and Rh. haemaphysaloides collected from stray animals in Taiwan could be infected the zoonotic pathogen R. felis. Moreover, R. rhipicephali was only identified in adult stage of Rhipicephalus sanguineus and Rh. haemaphysaloides.     

Isolation and Characterization of Large Marine Bacteriophage (Myoviridae), VhKM4 Infecting Vibrio harveyi

Abstract

The causative agent responsible for vibriosis in tropical fish aquaculture, Vibrio harveyi, has become a major bacterial pathogen. Studies suggest that this bacterium has developed resistance to antibiotics commonly used in aquaculture. In view of this situation and the requirement for the proposed postantibiotic era, bacteriophage therapy seems to be a promising control strategy for fish vibriosis. In this study, a lytic Vibrio phage VhKM4 belonging to a member of large, marine Myoviridae was successfully isolated. It exhibited bacteriolysis to both V. harveyi VHJR7 and V. parahaemolyticus ATCC 17802. The latent period of the VhKM4 phage was recorded at 60 min. It also recorded average burst size of approximately 52 plaque-forming units per infected cell. A strong bacteriolytic activity at low multiplicity of infection of 0.01 indicates the effectiveness of this large marine myovirid against fish pathogenic strain of V. harveyi VHJR7.

Received June 16, 2016; accepted October 7, 2016 Published online February 6, 2017     

Studies on synchronous egress of coccidian parasites (<Emphasis Type="Italic">Neospora caninum,Toxoplasma gondii,Eimeria bovis)</Emphasis> from bovine endothelial host cells mediated by calcium ionophore A23187

Neospora caninum, Toxoplasma gondii and Eimeria bovis are coccidian parasites of veterinary importance. Tachyzoites of N. caninum and T. gondii and sporozoites of E. bovis are able to invade and replicate in endothelial cells in vivo and in vitro. As it holds true for all eukaryotic cells, the survival of parasitized host cells and the parasites themselves should be dependent on ion balances, especially on extra- and intracellular calcium concentrations. Addition of the calcium ionophore A23187 reliably did release merozoites from mature N. caninum and T. gondii meronts grown in cultured primary bovine umbilical vein endothelial cells (BUVEC). Extent and time course of merozoite release depended on both, maturity of the meronts and concentration of the calcium ionophore. Attempts, however, to achieve synchronous release of merozoites from E. bovis first generation meronts by ionophore treatment failed, suggesting a different biological behaviour of this parasite. According to microscopical observations, the quite variable time of E. bovis macromeront maturation and a hampered merozoite exit owing to dense parasite-induced cytoskeleton elements surrounding the meront may be a reason for the lack of inducible synchronous release. Electronic supplementary materials The online version of this article (doi: ) contains supplementary material, which is available to authorized users.     

Up‐Regulation of Insulin‐Like Growth Factor I and Uteroglobin in In Vivo‐Developed Parthenogenetic Embryos

Parthenote embryos are being considered as an alternative source of embryonic stem cells. However, as there is still a dearth of knowledge of this kind of embryos, a better understanding of their biology is needed for their application. In this work, we studied the differences and similarities between parthenotes and normal embryos at the blastocyst stage in vivo developed. We analysed the expression of factor OCT‐4, vascular endothelial growth factor (VEGF), insulin‐like growth factor I (IGF‐I) and uteroglobin (UG) by real‐time PCR. To do so, oocytes were recovered and after activation procedure were transferred by ventral middle laparoscopy to receptive does to undergo completely in vivo development. Does were slaughtered 6 days post‐ovulation induction, and parthenote and normal embryos were recovered for mRNA expression analysis. Our results reported that parthenotes and normal embryos showed similar mRNA expression for OCT‐4 and VEGF. However, IGF‐I and UG showed to be over‐expressed in parthenote embryos. Thus, our study highlights that despite the in vivo development of parthenotes, they still seem to have an altered expression and, therefore, to be different to normal embryos. The altered expression pattern of parthenote embryos suggests that these embryos should be studied carefully before future application.     

Investigation of bovine brucellosis in the Northeastern Turkey

Bovine brucellosis, caused by Brucella abortus, is a significant problem for both public and animal health in Turkey. This study was conducted on the calving seasons between 2001 and 2006. A total of 626 serum samples of cattle obtained from 27 herds with a history of abortions was examined for Brucella antibodies by RBPT, SAT and ELISA. Of the cattle sera analysed, 221 (35,30%) and 206 (32, 92%) and 247 (39,45%) were found to be positive by RBPT , SAT and ELISA, respectively. B. abortus was isolated from 48 (32,21%) of 149 lung samples and stomach contents of the aborted fetuses. Based on the biochemical tests and the agglutination tests with monospecific A and M antisera, only 3 of the isolates were found to be B. abortus biotype 1 and the remaining 45 were biotype 3. This study also revealed that the dominant biotype of B. abortus was biotype 3 in this region. The determination of the agents responsible for bovine brucellosis and serosurvey of this disease are expected to help better understanding of this zoonotic infection in this region and neighbouring countries.     

An initial coprological survey of parasitic fauna in the wild Amur leopard (Panthera pardus orientalis)

The Amur leopard, one of nine recently recognized subspecies of leopard, is still the most threatened by a stochastic procession of extinction. Evaluation of the potential danger to the conservation of the Amur leopard originating from disease urgently needs to be studied. Unfortunately, research on the potential risk to Amur leopards caused by disease is rare. In terms of parasitic diseases that affect this species, even basic data for parasitic fauna are absent. The aim of this study is to acquire this knowledge to improve the general understanding of Amur leopard parasites. Seven parasite species, including 3 nematodes (Toxocara cati, a capillarid‐type parasite, and a Metastrongyloidea‐type parasite), 2 cestodes (Spirometra sp. and Taenia sp.), 1 trematode (Paragonimus sp.), and 1 protozoan (Cystoisospora felis), were found in this research. Toxocara cati occurred most frequently, followed by Spirometra sp.     

Inclusion keratoconjunctivitis ('pink eye') in sheep

Summary

The cytoplasnwtic inclusion bodies, which, in 1931. Coles discovered in the corneal cet. of sheep suffering from contagious keratoconjunctivitis arc now considered to be the reticulate bodies of a chlamydia. Colesiota conjunctivae (synonym: Chlamydia psittaci ovis).

According to the postulates of Koch Colesiota conjunctivae is a primary cause of contagious kerato‐conjunctivitis in sheep, but the clinical picture is complex and is a result of the interaction between the infecting chlamydiae. host resistance factors. and secondary infections caused by opportunistic bacterial ocular pathogens.

The clinical syndrome might also be caused by other micro‐organisms, such as Mycoplasma conjunctivae or environmental factors, such as dust. However, in these cases, cytopiasnwtic inclusion bodies cannot be found in the corneal cells of diseased eyes.

To differentiate chlamydial keratoconjunctivitis from keratoconjunctivitis due to other causes, it is proposed to include in the name the laboratory findings typical for this disease: Sheep Inclusion Keratoconjunctivitis.

Chlamydia are Gram‐negative bacteria, which arc obligate intracellular parasites.

Prolonged treatment seems to be required to eradicate chlamydiae from a host and antibiotics must reach intracellular levels that are higher than their minimum inhibitory concentration for chlamydiae. Tetracyclines are the drugs of choice. This means that for a microbiological cure, diseased sheep must be injected several times a day for a week or more. Because the disease is usually self‐limiting and economic losses are considered low, this seems unnecessary and control of the disease by local treatment of secondary infections .seems sufficient.

However, this will not prevent .spreading of the disease in a herd and relapses may occur.     

Distribution of angiostrongylosis in Cornwall

One hundred and ninety-seven faecal samples were examined for Angiostrongylus vasorum larvae, from dogs throughout Cornwall, which were presented with one or more clinical signs consistent with infection, were known to eat slugs or snails or were related to an infected dog. Twenty samples (10-1 per cent) contained nematode larvae, of these A vasorum larvae were identified in eight (4-1 per cent) and larvae of Filaroides species in eight (4-1 per cent). Seven of the eight dogs positive for A vasorum were found to have been infected within an area six miles in diameter incorporating Redruth. Statistical analysis suggested that this aggregation of cases was unlikely to be a random event (P < 0–01). No correlation between published data on the distribution of the intermediate host, slugs and snails and the cases of A vasorum infection reported in this paper could be found. It is concluded that A vasorum infection is enzootic within a small area around Redruth in the mid-west of Cornwall.     

Methicillin and aminoglycoside resistance in <Emphasis Type="Italic">Staphylococcus aureus</Emphasis> isolates from bovine mastitis and sequence analysis of their <Emphasis Type="Italic">mecA</Emphasis> genes

Although methicillin-resistant Staphylococcus aureus (MRSA) were generally isolated from human beings; these agents were recently isolated from various animal species. It has been shown that MRSA isolates are not only resistant to beta-lactam antibiotics, but can also be resistant to the other commonly used antibiotics. In this study, 18 phenotypic methicillin resistant S. aureus isolates from bovine mastitis cases were analyzed by PCR for the presence of mecA gene encoding methicillin resistance and aac(6′)/aph(2″), aph(3′)-IIIa and ant(4′)-Ia genes encoding aminoglycoside resistance. Out of 18 S. aureus isolates (oxacillin MICs, ≥4 μg/ml), 3 were positive for mecA gene. Only one from 3 mecA positive isolates was positive for genes encoding aminoglycoside-modifying enzymes and this isolate carried aac(6′)/aph(2″) in combination with aph(3′)-IIIa gene. The aph(3′)-IIIa gene was detected in 3 isolates. These three isolates carrying the aminoglycoside-modifying enzyme genes were resistant to gentamicin, kanamycin and neomycin. The mecA gene of 3 MRSA isolates was sequenced. All three mecA genes of these isolates were identical to that found in human MRSA strains, except a one-base substitution at nucleotide position 757. From the data presented in this study, it can be concluded that MRSA isolated from bovine mastitis may be originated from human beings, but further studies are needed to investigate the possibility of zoonotic transfer of MRSA.     

Leptospira interrogans at the Human–Wildlife Interface in Northern Botswana: A Newly Identified Public Health Threat

Leptospirosis is the most widespread zoonosis in the world. In northern Botswana, humans live in close proximity to a diversity of wildlife and peridomestic rodents and may be exposed to a variety of zoonotic pathogens. Little is known regarding the occurrence and epidemiology of L. interrogans in Africa despite the recognized global importance of this zoonotic disease and the threat it poses to public health. In Botswana, banded mongooses (Mungos mungo) live in close proximity to humans across protected and unprotected landscapes and may be a useful sentinel species for assessing the occurrence of zoonotic organisms, such as L. interrogans. We utilized PCR to screen banded mongoose kidneys for leptospiral DNA and identified 41.5% prevalence of renal carriage of L. interrogans (exact binomial 95% CI 27.7–56.7%, n = 41). Renal carriage was also detected in one Selous' mongoose (Paracynictis selousi). This is the first published confirmation of carriage of L. interrogans in either species. This is also the first report of L. interrogans occurrence in northern Botswana and the only report of this organism in a wildlife host in the country. Pathogenic Leptospira are usually transmitted indirectly to humans through soil or water contaminated with infected urine. Other avenues, such as direct contact between humans and wildlife, as well as consumption of mongooses and other wildlife as bushmeat, may pose additional exposure risk and must be considered in public health management of this newly identified zoonotic disease threat. There is a critical need to characterize host species involvement and pathogen transmission dynamics, including human–wildlife interactions that may increase human exposure potential and infection risk. We recommend that public health strategy be modified to include sensitization of medical practitioners to the presence of L. interrogans in the region, the potential for human infection, and implementation of clinical screening. This study illustrates the need for increased focus on neglected zoonotic diseases as they present an important threat to public health.