Esraa A. Elshafiee, Sara M. Nader, Sohad M. Dorgham and Dalia A. Hamza
Introduction: Carbapenem-resistant Pseudomonas aeruginosa (CRPA) has become the leading cause of health care-associated infections. Treatment is difficult due to the lack of an effective antimicrobial therapy, and mortality is high. This study investigated the occurrence of CRPA in farm animals (buffaloes and cattle), livestock drinking water, and humans in Egypt.
Material and Methods: A total of 180 samples were examined: 50 faecal each from buffaloes and cattle, 30 of livestock drinking water, and 50 stool from humans. The samples were cultured on cetrimide agar and the plates were incubated aerobically at 37°C for 24 h. The isolates were examined for the presence of the blaKPC, blaOXA-48, and blaNDM carbapenemase-encoding genes using PCR and investigated for the exotoxin A (toxA) gene. The toxA gene from carbapenem-group resistant isolates was phylogenetically analysed.
Results:P. aeruginosa was isolated from buffaloes, cattle, drinking water, and humans, with occurrences of 40%, 34%, 10%, and 20%, respectively. Carbapenem resistance genes were found in 60%, 59%, 67%, and 70% in buffalo, cattle, water and human samples, respectively. The toxA gene was detected in 80% of samples. The phylogenetic analysis showed that cattle and water sequences were in one cluster and more related to each other than to human isolates.
Conclusion: Occurrence of CRPA among farm animals, drinking water, and humans was high, reflecting the environmental origin of P. aeruginosa and highlighting contaminated water as a potential transmitter of CRPA to livestock and next to humans.
Jia-San Zheng, Jing-Nie, Ting-Ting Zhu, Hong-Ri Ruan, Xue-Wei and Rui-Wu
Introduction: The value of neutrophil gelatinase-associated lipocalin (NGAL), kidney injury molecule-1 (Kim-1), and liver-type fatty acid binding protein (L-FABP) was assessed in early diagnosis of gentamicin-induced acute kidney injury (AKI) in dogs.
Material and Methods: Subcutaneous gentamicin injection in 16 healthy adult beagles made the AKI model. Blood was sampled every 6 h to detect NGAL, Kim-1, L-FABP, and serum creatinine (SCr) concentrations. Kidney tissue of two dogs was taken before the injection, as soon as SCr was elevated (78 μmol/L), and when it had risen to 1.5 times the baseline, and haematoxylin-eosin staining and transmission electron microscopy (TEM) were used to observe changes.
Results: NGAL, Kim-1, and SCr levels were significantly increased (P < 0.05) at 18, 30, and 78 h post injection, but L-FABP concentration was not associated with renal injury. At the earliest SCr elevation stage, findings were mild oedema, degeneration, and vacuolisation in renal tubular epithelial cells in pathology, and mild cytoplasmic and mitochondrial oedema in TEM. At this time point, NGAL and Kim-1 concentrations were significantly increased (P < 0.05), indicating that these two molecules biomark early kidney injury in dogs. Using receiver operating characteristic curve analysis, their warning levels were > 25.31 ng/mL and > 48.52 pg/mL.
Conclusion: Plasma NGAL and Kim-1 above warning levels are early indicators of gentamicin-induced AKI in dogs.
Joanna Kochan, Wojciech Niżański, Nei Moreira, Zalmir Cubas da Silva, Agnieszka Nowak, Sylwia Prochowska, Agnieszka Partyka, Wiesława Młodawska and Józef Skotnicki
With the exception of the domestic cat, all felid species (Felidae) are currently threatened with extinction in their natural habitat. To develop effective and optimal wild cat conservation programmes with assisted reproductive technology (ART) it is necessary to combine advances from different disciplines of science, starting from the biology of the species, through research into the population and habitat, assisted reproductive technologies, establishment of gene banks, developing bioinformatic systems, and ending with biodiversity and endangered species management. In the last few years knowledge of felid reproduction has expanded considerably thanks to comparative studies utilising the domestic cat as a research model for endangered wild cats. Basic reproductive techniques utilised in both domestic cat breeding and rescuing wild felid populations that are threatened with extinction include semen collection and cryopreservation, artificial insemination, oocyte collection, in vitro maturation, in vitro fertilisation, somatic cloning, and embryo transfer. The main directions in which assisted reproductive technologies are being developed in wild cat conservation implementations and the contribution of Polish research centres in advancing these methods are presented.
Anna Grzesiakowska, Przemysław Baran, Marta Kuchta-Gładysz and Olga Szeleszczuk
Introduction: Comparative analysis of the karyotype structure was made in two hedgehog species: the northern white-breasted hedgehog (Erinaceus roumanicus) and the African pygmy hedgehog (Atelerix albiventris). Material and Methods: The cytogenetic analysis used differential staining techniques (DAPI, Ag-NOR, and C-banding/DAPI) and sequential QFQ/FISH banding with NOR20 and TEL20 probes which showed 45S rDNA and (TTAGGG)n repeat sequences, respectively, on hedgehog chromosomes. Results: It was confirmed that the somatic cells of the hedgehogs have a constant number of chromosomes (2n = 48,XY). Differences were observed in the NOR number between the species. NORs were identified on three autosome pairs in the northern white-breasted hedgehog and on only two pairs in the African pygmy hedgehog. Chromosome analysis by C-banding/DAPI showed large segments of heterochromatin rich in A-T pairs on three autosome pairs in both the northern white-breasted and African pygmy hedgehogs. The heterochromatin segments encompassed large fragments of the longer arm of chromosome pairs 13, 14 and 20. The (TTAGGG)n repeat sequences on the hedgehog chromosomes were only observed in the terminal position of all the chromosomes in both species. Conclusion: Our observations provide new information on the level of diversity within the Erinaceidae family.
Yu-Xi Song, Pan Hu, Yun-long Bai, Chang Zhao, Cheng Xia and Chuang Xu
Introduction: Differential metabolites (DMs) between cows with inactive ovaries (IO) and oestrous (E) cows were screened and metabolic pathways of DMs associated with IO were determined. Material and Methods: Cows at 50 to 60 days (d) postpartum from an intensive dairy farm were randomly selected and allocated into an E group (n = 16) or an IO group (n = 16) according to a pedometer and rectal examinations. Their plasma samples were analysed by liquid chromatography–mass spectrometry (LC–MS) to compare plasma metabolic changes between the E and IO groups. Multivariate pattern recognition was used to screen the DMs in the plasma of IO cows. Results: Compared with normal E cows, there were abnormalities in 20 metabolites in IO cows, including a significantly decreased content (VIP > 1, P < 0.05) of cholic acid, p-chlorophenylalanine, and arachidonic acid, and a significantly increased content (VIP > 1, P < 0.05) of tyramine, betaine, L-phenylalanine, L-glutamate, D-proline, L-alanine, and L-pyrophosphate. Five DMs (cholic acid, D-proline, L-glutamate, L-alanine, and L-pyroglutamic acid) with higher variable importance in projection (VIP) values between groups were validated by ELISA with blind samples of re-selected cows (IO, 50 to 60 d postpartum) and the validated results were consistent with the LC–MS results. Conclusion: The 20 DMs in IO cows during the peak of lactation indicated that the pathogenesis of IO was involved in complex metabolic networks and signal transduction pathways. This study provides a basis for further exploration of the pathogenesis and prevention of IO in cows in the future.
Jim Weber, Markus Zenker, Gábor Köller, Manfred Fürll and Markus Freick
Introduction: Recumbency is a frequent symptom occurring throughout lactation. Its cause can be related to the energy or mineral metabolism, or to trauma or infectious diseases. We compared various clinical chemistry parameters between healthy and recumbent cows and between cows with different causes of recumbency and determined if hypocalcaemia manifests in later lactation. Material and Methods: Recumbent (n = 32) and healthy (n = 32) German Holstein cows were studied. After clinical examination, a serum sample was taken to measure the concentrations of Mg, Ca, Fe, Na, K, Pi, β-hydroxybutyrate, total bilirubin, non-esterified fatty acids (NEFA), urea, and creatinine as well as activities of alkaline phosphatase, aspartate aminotransferase (AST), creatine kinase (CK), and γ-glutamyl transferase in recumbent cows > 5 d in milk and control cows matched for age, lactation number, and pregnancy stage. Results: In recumbent cows, mean serum concentrations of NEFA, bilirubin, and CK were statistically higher, while those of Fe, K, and Pi were significantly lower. Parameters compared between different recumbency diagnoses showed some descriptive Fe, K, urea, and AST differences, but these were not statistically significant. Conclusion: The results show that only a limited number of parameters have diagnostic besides therapeutic value. Although of minor importance in our study, hypocalcaemia should be considered a cause of recumbency, even outside the typical risk period of parturient paresis.
Konrad Pietruk, Marta Piątkowska and Małgorzata Olejnik
Introduction: Some azo dyes, including Sudans I–IV and Para Red, are genotoxic and may be biotransformed to cancerogenic aromatic amines. They are banned as food and feed additives, but their presence has been detected in food. Aromatic amines are also considered potentially toxic. Online EC–MS is a promising tool to study the transformation mechanisms of xenobiotics such as azo dyes. The aim of the study was to investigate emulation of how azo dyes are enzymatically transformed to amines with EC–MS. Material and Methods: The reduction reactions of five azo-dyes (Sudans I–IV and Para Red) were conducted using a glassy carbon working electrode and 0.1% formic acid in acetonitrile. Reduction results were compared with the literature and in silico to select preliminary candidates for metabolites. The LC-MS/MS method was used to confirm results obtained by electrochemical reactor. Results: A limited number of pre-selected compounds were confirmed as azo-dyes metabolites – aniline for Sudan I, aniline and 4-aminoazobenzene for Sudan III, o-toluidine for Sudan IV, and 4-nitroaniline for Para Red. No metabolites were found for Sudan II. Conclusions: Electrochemistry–mass spectrometry was successfully applied to azo-dyes. This approach may be used to mimic the metabolism of azo-dyes, and therefore predict products of biotransformation.
Olga Gójska-Zygner, Justyna Bartosik, Paweł Górski and Wojciech Zygner
Introduction: A previous study on canine babesiosis showed low serum tonicity in affected dogs, which may result from syndrome of inappropriate antidiuretic hormone secretion (SIADH). This endocrine disorder was recognised in human malaria which is considered a disease with similar pathogenesis to canine babesiosis. The aim of this study was to investigate the occurrence of SIADH in babesiosis-afflicted dogs. Material and Methods: Serum and urinary sodium and urine specific gravity (USG) were determined in dogs with babesiosis. Mean arterial pressure (MAP) was measured at the beginning of the clinical examination. Serum tonicity and osmolality were calculated. Correlations were calculated between MAP and serum and urinary sodium concentrations, USG, serum tonicity, and calculated serum osmolality. Results: Statistically significant correlations were observed between MAP and tonicity, calculated osmolality, USG, and serum and urinary sodium concentrations in non-azotaemic dogs. In three non-azotaemic dogs SIADH was recognised. Conclusion: SIADH develops in non-azotaemic dogs with babesiosis. It is probably associated with decreased blood pressure in infected dogs. Thus, it seems that in fact it may be appropriate vasopressin secretion in canine babesiosis as a protective mechanism in hypotension which leads to hypoxia and renal failure in affected dogs.
Hatice Esra Çolakoğlu, Murat Onur Yazlık, Mert Pekcan, Ufuk Kaya, Cihan Kaçar, Mehmet Rıfat Vural, Serdal Kurt, Merve Menekse Yildirim, Ahmet Bas and Şükrü Küplülü
Introduction: The objectives of this study were to determine the role of a fall in pre-calving body condition score (BCS) in postpartum metabolic status and reproductive outcomes, and gauge the indicativeness of blood metabolites during the transition period. Material and Methods: Cows were grouped based on BCS loss between days −14 ±3 and 0 relative to calving. Cows that lost no BCS were the BCS control group (BCS-C), cows that lost 0.25 BCS points the low BCS loss group (BCS-L), and those that lost 0.5 points or more the high BCS loss (BCS-H) group. Blood was taken on days −14 ±3, 3, 14, and 30 ±4 for determination of comprehensive metabolic panel biomarker levels. Beta-hydroxybutyric acid (BHBA) levels were quantified on postpartum examination days. Vaginal discharge scores, ovarian activity on day 30 ±4, and subsequent fertility parameters were evaluated. Results: The BCS-H cows had lower mean Ca concentrations before calving and on day 3, when the group’s BHBA and CK were higher (P < 0.05); on day 14 they had higher AST concentrations (P < 0.05). The BCS-L cows had greater bilirubin levels (P < 0.05). The BCS-H cows had lower cyclicity and higher endometritis rates. First service pregnancy rates were 50%, 50%, and 61.9%, open days 96.8, 95.75, and 89.2, and overall pregnancy rates 56.25%, 65%, and 80.95 % in the BCS-H, BCS-L, and BCS-C groups, respectively. Conclusion: Prepartum BCS loss of ≥ 0.5 points could be associated with Brown Swiss cow low Ca and BHBA concentrations early postpartum, and with subsequent uterine health and overall pregnancy rate. Prepartum Ca concentration might be a prognostic biomarker for postpartum metabolic status and reproductive outcomes.
Epy Muhammad Luqman, I Ketut Sudiana, Win Darmanto, Agung Budianto Achmad and Widjiati
Introduction: The aim of the study was to describe the process of neuron death in the cerebral cortex caused by embryonic carbofuran exposure. Material and Methods: 81 mouse foetuses from 27 breeding mice were used in the study. Carbofuran was administered by gavage from the 6th to the 15th day of gestation to two groups: one at 0.0208 and the other at 0.0417 mg/kg b.w. On the 17th day, the mice were sacrificed and the foetuses were taken to measure the ROS (malondialdehyde/MDA and superoxide dismutase/SOD) activity in brain tissue, the number of apoptotic embryonic cerebral cortex neurons using a TUNEL assay, and necrotic cells using HE staining. Examination of p53 and caspase 3 expression was done by immunohistochemistry. Data were analysed using analysis of variance (ANOVA) and Duncan’s test. Results: Increased activity of cerebral ROS characterised by significant elevation of the MDA level (P < 0.05), decreased SOD (P < 0.01), increased p53 and caspase 3 expression, and cerebral cortical neuron death either by necrosis or apoptosis (P < 0.05) were found. At the low dose carbofuran increased expression of p53, caspase 3, and apoptosis. At the high dose it increased levels of MDA and necrosis. Conclusion: Increased expression of p53 and caspase 3 and apoptosis indicated that carbofuran may cause apoptosis through the intrinsic pathway. The increased apoptosis grants an opportunity to prevent and treat the effect of ROS due to gestational carbofuran exposure.