The proboscis monkey (Nasalis larvatus) is an endangered species with a declining population. This article describes the first successful attempt at sperm collection and evaluation, and the testicular and sperm morphometries of the wild proboscis monkey in Sabah, Malaysia. Eight semen collection procedures using electro-ejaculation and digital manipulation were conducted in three wild adult male proboscis monkeys. A total of 21 ejaculates were collected. The testicular biometry was measured with the aid of ultrasonography. Sample evaluation included semen volume and pH and sperm concentration, viability, and abnormality. The sperm morphometry was undertaken using phase contrast microscopy. The mean (±SD) total testicular volume of these animals was 5.77 cm³ (±1.58). Semen collection by electro-ejaculation resulted in an 84% success rate, while digital manipulation did not result in any ejaculation. Each animal showed different semen characteristics, where the volume was 5–540 μL, pH 8–9, and sperm concentration 0.041–83.00 ×106/mL. The percentage of abnormal sperm was high at 76.8% (±89.60), largely due to midpiece abnormality. Normal sperm had a spherical head and long tail with a head : midpiece : tail length ratio of 1 : 2: 8. The social status of these animals may contribute to the generally low quality of the semen. The techniques and data from this study are useful for future conservation and application of assisted reproductive technology in this species.

The proboscis monkey (Nasalis larvatus) is an endangered species with a declining population. This article describes the first successful attempt at sperm collection and evaluation, and the testicular and sperm morphometries of the wild proboscis monkey in Sabah, Malaysia.

The objective of this study was to determine the prevalence and characteristics of antimicrobial-resistant Enterococcus faecalis and E. faecium isolated from the oral cavities of captive giant pandas in China. The virulence-associated determinant and antimicrobial resistance genes were detected and antimicrobial susceptibility tests were performed on 54 strains of each bacterium. All isolates showed 100% multidrug resistance. E. faecalis isolates showed a higher percentage of strains resistant to gentamicin (48.1%), vancomycin (55.6%), linezolid (100%), and streptomycin (33.3%) than E. faecium isolates. The resistance genes of Enterococcus spp. were present to highly varying extents according to antibiotic type, their presence breaking down for E. faecalis and E. faecium respectively as aac(6')/aph(2″) 5.56% and 5.56%; aph(3')-Ⅲ 0% and 14.81%; ant(6)-I 0% and 3.7%; ant(4')-Ia 0% and 64.81%; tetL 20.37% and 100%; vanA 92.59% and 46.3%; vanB 0% and 0%; cfr 0% and 90.74%; optrA 96.3% and 3.7%; blaZ 0% and 1.85%; blaTEM 0% and 0%; tetA 20.37% and 0%; tetC 24.07% and 100%; tetM 0% and 0%; ermA 12.96% and 100%; ermB 5.56% and 3.7%; and ermC 0% and 1.85%.Virulence-associated determinants were detected in this research, which typically include efaA, gelE, asa1, ace, cylA, esp and hyl; however, the latter three were not detected. High proportions of the isolates carried the efaA, gelE, asa1, and ace genes. Respectively for E. faecalis and E. faecium their detection was efaA 98.1% and 85.2%; gelE 98.1% and 87%; asa1 92.6% and 87%; and ace 87% and 85.2%. This is the first study on the potential disease risk and antimicrobial-resistant characteristics of E. faecalis and E. faecium isolates in giant panda oral cavities. The results of this study show that the antimicrobial resistance rate of Enterococcus spp. isolated from the oral cavity of captive pandas is very high, and thus needs to be monitored.

The objective of this study was to determine the prevalence and characteristics of antimicrobial-resistant Enterococcus faecalis and E. faecium isolated from the oral cavities of captive giant pandas in China.

H7N9 avian influenza has broken out in Chinese poultry 10 times since 2013 and impacted the industry severely. Although the epidemic is currently under control, there is still a latent threat. Epidemiological surveillance data for non-human H7N9 avian influenza from April 2013 to April 2020 were used to analyse the regional distribution and spatial correlations of positivity rates in different months and years and before and after comprehensive immunisation. In addition, positivity rate monitoring data were disaggregated into a low-frequency and a high-frequency trend sequence by wavelet packet decomposition (WPD). The particle swarm optimisation algorithm was adopted to optimise the least squares support-vector machine (LS-SVM) model parameters to predict the low-frequency trend sequence, and the autoregressive integrated moving average (ARIMA) model was used to predict the high-frequency one. Ultimately, an LS-SVM-ARIMA combined model based on WPD was constructed. The virus positivity rate was the highest in late spring and early summer, and overall it fell significantly after comprehensive immunisation. Except for the year 2015 and the single month of December from 2013 to 2020, there was no significant spatiotemporal clustering in cumulative non-human H7N9 avian influenza virus detections. Compared with the ARIMA and LS-SVM models, the LS-SVM-ARIMA combined model based on WPD had the highest prediction accuracy. The mean absolute and root mean square errors were 2.4% and 2.0%, respectively. Low error measures prove the validity of this new prediction method and the combined model could be used for inference of future H7N9 avian influenza virus cases. Live poultry markets should be closed in late spring and early summer, and comprehensive H7N9 immunisation continued.

H7N9 avian influenza has broken out in Chinese poultry 10 times since 2013 and impacted the industry severely. Although the epidemic is currently under control, there is still a latent threat.

Proper conformational arrangement of the E2 molecules of bovine viral diarrhoea-mucosal disease virus (BVD-MDV) is crucial to obtain an effective recombinant vaccine candidate against the disease. In this study, we characterised a new molecule composed of two distinct sequences of the E2 glycoprotein of BVD-MDV and the Fc fragment of human immunoglobulin (BVDE2Fc). The chimaeric protein was expressed in mammalian cell lines of different species by adenoviral transduction and purified by immobilised metal-affinity chromatography. The N-glycans were profiled by HPLC, and the BVDE2Fc immunogenicity was assessed in male mice. The antigen-antibody reactions were evaluated by ELISA. The MDBK cell line was selected from among five for the final production of BVDE2Fc. After purification to over 90%, the N-glycan profile showed neutral and complex oligosaccharides. The mouse immunisation induced a strong humoral response, which produced antibodies able to attach to conformational epitopes on E2 molecules, while the Fc fragment barely contributed to the immune response. Additionally, BVDE2Fc attached to antibodies from bovine sera positive to distinct BVD-MDV subtypes, whereas the loss of BVDE2Fc structure during the deglycosylation process considerably diminished those interactions. These results demonstrate that the structure of E2 molecules arranged in tandem and attached to an Fc fragment could represent a viable design for future vaccine candidates against BVD-MD.

Proper conformational arrangement of the E2 molecules of bovine viral diarrhoea-mucosal disease virus (BVD-MDV) is crucial to obtain an effective recombinant vaccine candidate against the disease. In this study, we characterised a new molecule composed of two distinct sequences of the E2 glycoprotein of BVD-MDV and the Fc fragment of human immunoglobulin (BVDE2Fc).

Globally, genetically modified (GM) crops were grown on 191.7 million hectares in 2018, which were mostly sown with soybean, maize, cotton, oilseed rape, and rice. The most popular traits introduced through genetic modification include herbicide and pest insect resistance. The aim of this study was to identify and quantify genetically modified soybean used in animal feed in Poland. This research was based on the real-time PCR technique. All methods for GM soybean events were adopted from the EURL GMFF database of methods and previously verified to meet the minimum criteria of acceptance. Over 15 years of research, 665 samples were examined in total. The most common GM soybean event was MON40-3-2, tested for from the beginning of the investigation. Next, in decreasing order of frequency, were MON89788, MON87701, and A2704-12. In the majority of samples (606; 91%) GM soybeans were identified at a content level above the 0.9% GM content threshold for mandatory labelling. Only 59 soybean samples (9%) were identified as GM negative. GM negative results were mainly identified during the analyses in the last three years of the study, from 2017 to 2019. Our data clearly indicate that the majority of soybean used in Poland for animal feeding was genetically modified.

Globally, genetically modified (GM) crops were grown on 191.7 million hectares in 2018, which were mostly sown with soybean, maize, cotton, oilseed rape, and rice. The most popular traits introduced through genetic modification include herbicide and pest insect resistance. The aim of this study was to identify and quantify genetically modified soybean used in animal feed in Poland.

The aim of the study was to determine how the spread of contagious agalactia in sheep and goats in the Odesa region depended on the age of the animals and the season. From January 2016 to December 2018, 1,964 ewes and 1,484 nanny goats of different age groups were studied by ELISA for antibodies to Mycoplasma agalactiae. The highest incidence of contagious agalactia was registered in one-year-old animals and was 59.7‒83.0%, two-year-old ruminants showed 17.0‒40.3% prevalence, in livestock at the age of 3–4 years no serological evidence of the disease was registered and in ewes and nanny goats older than 5–6 years 1.5–3.6% were infected. The most susceptible were young animals at the age of one-month (11.6‒14.5%). The first peak of the disease was recorded in March‒April (21.0‒26.1%), in the lambing period, which coincided with the beginning of lactation and the suckling period, and the second peak occurred in June–July (28.9‒34.2%), the period of maximum lactation and of manual milking of sheep and goats. The results of serological investigations indicate the circulation of M. agalactiae in small ruminants in the south of Ukraine. To avoid greater dissemination of the pathogen, appropriate measures should be applied and strategies for its control need to be drawn up.

The aim of the study was to determine how the spread of contagious agalactia in sheep and goats in the Odesa region depended on the age of the animals and the season.

The aim of this study was to evaluate the effect of negative energy balance (NEB) on the final growth of the dominant ovarian follicle in Holstein cows. Cows at 14 to 21 d postpartum from an intensive dairy farm were randomly selected and allocated into a positive energy balance group (PEB, with β-hydroxybutyric acid (BHBA) level < 1.2 mmol/L, n = 15) and an NEB group (BHBA > 1.2 mmol/L, n = 15). Plasma samples were collected at 21, 50 and 55 d postpartum to assess the concentrations of energy metabolites, minerals, hormones and cytokines. Ovaries were examined by transrectal ultrasound on days 50 and 55 (120 hours later) to evaluate the diameter of the largest follicle. Compared with PEB cows, there were a more severe body condition loss and a lower milk yield in NEB cows (P < 0.05) and these had greater concentrations of plasma BHBA, non-esterified fatty acids, triglycerides, urea nitrogen, growth hormone, interleukin 6, and fibroblast growth factor 21 and lesser concentrations of plasma glucose, total cholesterol, insulin, insulin-like growth factor 1, insulin-like growth factor binding protein 3, leptin, brain-derived neurotrophic factor and angiopoietin-like protein 8 on d 21 (P < 0.05), while plasma minerals were not affected by energy status (P > 0.05). These changes persisted until the end of the study period (50–55 days postpartum) resulting in a lower follicular growth rate for cows in the NEB than the PEB group. These observations indicate that follicular growth rate is associated with measurable changes in energy metabolite, hormone and cytokine concentrations caused by early postpartum NEB.

The aim of this study was to evaluate the effect of negative energy balance (NEB) on the final growth of the dominant ovarian follicle in Holstein cows.

Apocrine sweat gland carcinomas (ASGCs) are rare malignant skin tumours in dogs and humans. The literature published so far focuses mostly on the clinico-epidemiological aspect of these tumours, but little is known about their pathogenesis. In this study we aimed to determine whether the p53 gene is involved in the carcinogenesis of the apocrine sweat gland in dogs and whether ultraviolet radiation (UV) is related to it. Forty canine ASGCs were submitted to laser capture microdissection to isolate neoplastic cells, from which DNA was subsequently extracted. PCR amplification and sequencing of p53 exons 2–8 was then performed, followed by computer analysis of the obtained sequences. Sixteen mutations within the p53 gene were found in 13 tumours. The mutations involved C → T, T → C, G → A, and CC → TT transitions, C → G transversion and adenine deletion, which are gene alteration types known to be related to UV radiation in the process of skin carcinogenesis in humans. Six of the thirteen tumour cases displayed the C → T transitions in the same location in exon 4 and three of the thirteen cases displayed T → C in the same location in exon 5. The results of the present study indicate both the participation of the p53 gene and the influence of UV radiation in the formation of ASGCs in dogs.

Apocrine sweat gland carcinomas (ASGCs) are rare malignant skin tumours in dogs and humans. The literature published so far focuses mostly on the clinico-epidemiological aspect of these tumours, but little is known about their pathogenesis. In this study we aimed to determine whether the p53 gene is involved in the carcinogenesis of the apocrine sweat gland in dogs and whether ultraviolet radiation (UV) is related to it.