Cerebral arterial vascularization of the scimitar-horned oryx (Oryx dammah).

The Scimitar-horned Oryx (Oryx dammah) is a large terrestrial mammal native to Africa. Since the year 2000, it is classified as extinct in the wild. It is a subject of various conservation projects. The aim of this study was to describe the arterial vascularization of the brain in this species of oryx. Three different anatomical methods were used to obtain a complete arterial pattern - latex injection, corrosion cast, and computed tomography. The arterial vascularization of the brain was described. The main components of the cerebral arterial circle were the rostral cerebral arteries and the caudal communicating arteries. These vessels were created from the intracranial part of the internal carotid artery, that emerged from the rostral epidural rete mirabile. In the juvenile specimen, the whole internal carotid artery was observed. The anatomical pathways of the blood supply to the brain are important during medical procedures in cases of congestion and fainting caused by inadequate brain blood perfusion.

Arterial Blood Supply to the Cerebral Arterial Circle in the Selected Species of Carnivora Order from Poland.

Carnivores are a wide, diverse group of mammals whose representatives live all over the world. The study presents the results of the analysis of the arterial vascularization of the blood supply to the cerebral arterial circle of selected species in the Caniformia suborder living in Poland. The selected group consists of wild and farm animals-105 animals in total. Three different methods were used-latex preparation, corrosion cast, and cone-beam computed tomography angiography. The main source of blood for encephalon in the described species is the internal carotid artery, and the second one is the vertebral artery. The results were discussed in relation to the current knowledge of this field of research. Information on the potential physiological meaning of such vascular pattern has been provided.

Cerebral Vascularization and the Remaining Area Supply of the Internal Carotid Artery Derivatives of the Red Kangaroo (Osphranter rufus).

The red kangaroo (Osphranter rufus) is a member of Macropidideae superfamily. It is one of the four kangaroo species living nowadays, and it is the biggest one. It is native to Australia, where it is an abundant species living across the whole continent in stable populations. Outside its natural habit, the red kangaroo is a common species found in zoos and as patients in wildlife rehabilitation centers. Reports on kangaroo anatomy are scarce. Describing detailed anatomy is a base for establishing diagnostic and treatment protocols for different species of animals. Cardiovascular diseases and pathological changes suggestive of hypertension have been previously described in kangaroos. This creates a necessity for detailed studies on species' vascular anatomy. New reports in the field of detailed vascular anatomy can bring considerable information that complements numerous studies on the evolution or biology of individual species. In this article, we describe the arterial vascularization of the brain and nearby regions of the cranial cavity using various anatomical techniques. The vascularization of the brain is discussed and compared with different mammalian species.

Cleft Lip and Palate in Four Full-Sib Puppies from a Single Litter of Staffordshire Bull Terrier Dogs: An Anatomical and Genetic Study.

Cleft lip and palate (CLP) is a well-known congenital defect in dogs, characterized by abnormal communication between the oral and nasal cavities. Its incidence rate is high and affects all dog breeds. The etiology of CLP is thought to be multifactorial, caused by both genetic and environmental factors. In this study, four puppies out of seven from a single litter of Staffordshire Bull Terrier dogs with craniofacial abnormalities were anatomically and genetically examined. Classical anatomical preparation, dyed-latex-injection of the arterial vessels, and cone-beam computed tomography were used. The puppies showed variations in their observable abnormalities: three of them had a complete cleft of the palate on both sides, while one puppy had a cleft on the right side only. Cytogenetic analysis showed a normal diploid chromosome number (2n = 78,XX or 78,XY) in the studied animals. Known genomic variants of CLP were examined in the ADAMTS20, DLX6, and MYH3 genes, but no mutations were identified. Further studies are needed to identify the breed-specific genetic variants associated with canine CLP.

Effects of raw and fermented rapeseed cake on ruminal fermentation, methane emission, and milk production in lactating dairy cows.

The objective of this study was to evaluate the effects of replacing raw rapeseed cake (RC) with fermented rapeseed cake (FRC) in the diet of dairy cows on methane (CH4) production, ruminal fermentation, and milk production, composition, and fatty acid composition. The Hohenheim gas test (exp. 1) was initially used to evaluate RC and FRC as substrates. Following batch fermentation, an in vitro study (exp. 2) was performed to assess the effects of replacing RC with FRC at 28.75, 57.5, 86.25, and 115 g/kg (FRC25, FRC50, FRC75, and FRC100) in the total mixed rations (TMR). Based on the in vitro results, the control TMR (115 g/kg dry matter (DM) of RC; CONRC) and experimental TMR (115 g/kg DM of FRC; FRC100) were chosen for an in vivo assessment. In exp. 3, four ruminally cannulated cows were used in a replicated 2 (group) × 2 (period) crossover design and fed the TMR ad libitum. In exp. 4, twenty multiparous Polish Holstein-Friesian cows in their mid-lactation (148 ± 26 d in milk) were used in a completely randomized design. The cows were fed a partial mixed ration without the RC and FRC, and the RC and FRC were supplied in a concentrate feeder at 2.65 kg/d/cow. The FRC100 markedly decreased CH4 production by 12% and archaeal population without adversely affecting nutrient digestibility. The molar proportion of propionate was increased, and the molar proportion of acetate and butyrate and acetate to propionate ratio were decreased by FRC100. No significant effects on milk production or composition, except an increase in milk urea concentration, were observed in cows fed FRC100. Milk C18:2 cis-9, trans-11 concentration was greater, and n-6 to n-3 fatty acid ratio was lower for FRC100 than CONRC. In-situ ruminal degradation of RC and FRC were explored using in-sacco techniques (exp. 5). The potential degradation and effective degradability of the DM, organic matter, and crude protein were significantly higher for FRC than RC. These results suggested that FRC could mitigate enteric CH4 production by decreasing archaeal abundances without adversely affecting milk production and ruminal fermentation in lactating cows.

Seroprevalence of Toxoplasma gondii in domestic cats, dogs and rabbits from Poland.

The seroprevalence of Toxoplasma gondii in domestic cats, dogs and rabbits was evaluated. Samples from cats and dogs were collected from five veterinary practices from various parts of Poland - Poznan (wielkopolskie voivodeship), Przemysl (podkarpackie voivodeship), Kluczbork (opolskie voivodeship), Lublin (lubelskie voivodeship) and Deblin (lubelskie voivodeship). Moreover, the samples from rabbits were collected in Poznan. In total, serum samples from 193 cats, 204 dogs and 71 rabbits were randomly selected and tested for specific antibodies against T. gondii using a commercial ELISA test. Pathogen seroprevalence among cats and dogs was calculated at a 95% confidence interval (CI) for each sex and age category (up to 12 months, 1-3 years, 4-7 years and over 8 years) and compared with a chi-squared test. The highest seroprevalence of T. gondii was noted in cats - 49.74% (96/193; 95% CI: 42.76-56.73). In dogs, it reached 28.92% (59/204; 95% CI: 23.13-35.49). Only 1 rabbit (3-year-old male) was seropositive, and the seroprevalence in rabbits was 1.41% (1/71; 95% CI: 0.25-7.56). A statistically significant correlation between seropositivity and age (p < 0.05) was observed in cats and dogs. No statistically significant difference in seroprevalence concerning gender or location was found in cats and dogs. Our findings indicate that cat and dog serum samples had a high frequency of anti-T. gondii antibodies, while rabbit serum samples had low frequency and that these species are exposed to T. gondii in Poland and develop humoral response due to infection.

Profile of Selected Mineral Elements in Tibiotarsal Bone of the White-Tailed Sea Eagle in Its Natural Habitat.

Mineral bone composition (dry matter, bones ash, P, Ca, Zn, Mn, Mg, and Cu) and Pb levels of tibiotarsi of seven White-Tailed Sea Eagles were assessed. Lead intoxication in different bird species including waterfowl and raptors is being studied worldwide. The bones were analyzed for Pb by mass spectrometry with excitation in inductively coupled plasma (ICP-MS Elan DRC II) and for bone composition by Atomic Emission Spectrometer (Agilent 4100 Microwave Plasma). Pb levels ranging from 3.54 µg/g to 74.6 µg/g DM suggest that some of the investigated birds might have been intoxicated by Pb. Results of this analysis were divided into two groups of bones, with bone Pb levels higher and lower than Pb toxicity levels, and mineral bone compositions of both groups were compared. The present study shows the differentiation of bone mineral composition among seven examined White-Tailed Sea Eagles, considered a specific species in raptors. Pb intoxication may not have a major influence on mineral bone composition in raptors. It also suggests that assessing bone composition of raptor bones may help finding the possible cause of their deaths.

Arterial Circle of the Brain of the Red-Necked Wallaby (Notamacropus rufogriseus).

The red-necked wallaby is a medium-sized marsupial species, which have increasingly been kept as pets around the world. In the study, the arterial blood supply for the brain in this species was described. The study was conducted on 50 specimens with two preparation methods. The main artery supplying the brain was the internal carotid artery. The arterial circle of the brain was closed from the caudal side. The anatomy of the arteries of the described region was compared with other groups of mammals. This is the first description of this anatomical area that has been carried out in a marsupial species. Understanding the anatomy of the circulatory system in the wallaby can be valuable for further physiological and pathophysiological studies.

Cytogenetic and molecular insight into the genetic background of disorders of sex development in seventeen cats.

The genetic background of feline disorders of sex development (DSDs) is poorly understood. We performed comprehensive cytogenetic, molecular, and histological studies of 17 cats with abnormal external genitalia, unusual behavior, or tricolor coats (atypical in males). The DSD phenotype of three cats was associated with sex chromosome abnormalities: X/Y translocation (38,XXSRY+), 37,X/38,XY mosaicism, and XX/XY leukocyte chimerism. The remaining 14 affected cats were classified as XY DSD (SRY-positive). In this group and 38 normal males, we analyzed a priori selected candidate genes (SRY, TAC3, CYP11B1 and LHCGR). Only a previously reported nonpathogenic variant was found in SRY. Moreover, SRY gene copy number was determined, and three variants were observed: 6, 5 (modal), and 4 copies in a single DSD case. The known variants in TAC3 and CYP11B1, responsible for testicular hypoplasia, persistent primary dentition or congenital adrenal hyperplasia, were not found in the study group. Nine novel polymorphisms were identified in the LHCGR gene, one of which, a potentially regulatory indel variant in 5'UTR, was significantly associated (p = 0.0467) with XY DSD. Our report confirmed that abnormalities of sex chromosomes are important causes of feline DSDs. We also showed that the indel variant of LHCGR can be considered a promising marker associated with XY DSD phenotype.

First detection of Hedgehog coronavirus 1 in Poland.

Hedgehogs are common in the majority of European countries and are known to host various pathogens, including viruses. The recent discovery of MERS-related coronaviruses (CoVs) in hedgehogs from Germany, France, the UK, China, and Italy suggests that hedgehogs may represent a wild reservoir of betacoronaviruses. This study reports the first detection and characterization of novel betacoronovirus, subgenus Merbecovirus in wild hedgehogs in Poland. The CoV RNA was detected in 10 out of 40 hedgehogs' rectal swabs and in 1 out of 18 samples of the lung. No viral RNA was identified in the duodenum and kidney. There was no significant relationship between clinical status, gender, hedgehogs' age, and coronaviral RNA detection. Phylogenetic analysis showed that CoVs detected in our study grouped together with other representatives of Hedgehog coronavirus 1 species identified in Western Europe. Our findings provide further evidence that hedgehogs are a natural reservoir of Merbecovirus. Considering the high mutation rate of CoVs and their potential for crossing interspecies barriers, the proper management of hedgehogs admitted to wildlife rehabilitation centres is needed. It cannot be excluded that merbecovirus strains detected in hedgehogs may recombine with other CoVs leading to new viruses with potential for interspecies transmission.

A cross-sectional retrospective study of SARS-CoV-2 seroprevalence in domestic cats, dogs and rabbits in Poland.

BACKGROUND: Coronaviruses (CoVs) have long been known to cause infection in domestic and free-living birds and mammals including humans. The zoonotic origin of SARS-CoV-2 and the biological properties of CoVs, including ability to cross interspecies barriers, enable its emergence in populations of various animals, including companion animals (cats, dogs, rabbits) an area requiring further study. To date, several cases of cats and dogs positive for SARS-CoV-2 and/or specific antibodies have been described. The aim of our cross-sectional retrospective study is to determine seroprevalence of SARS-CoV-2 in domestic dog, cat and rabbit population during recent COVID-19 pandemic in Poland. RESULTS: In total, serum samples from 279 cats and 343 dogs and 29 rabbits were used in the study. The seroprevalence of SARS-CoV-2 in cats and dogs reached 1.79% (95% CI: 0.77 - 4.13) and 1.17% (95% CI 0.45 - 2.96), respectively (p ≥ 0.05). Anti- SARS-CoV-2 antibodies were detected in 5 cats (mean S/P% 106 ± 48.23) and 4 dogs (mean S/P% 78.5 ± 16.58). All 29 samples from rabbits were negative for SARS-CoV-2 antibodies. No significant gender or age differences in seroprevalence in dogs and cats (p ≥ 0.05) were found. None of the animals with anti-SARS-CoV-2 antibodies displayed respiratory or gastrointestinal signs at the time of sampling. CONCLUSIONS: Our results confirmed previous findings that SARS-CoV-2 infections in companion animals occurs but are not frequent. Future serological testing of large pet population may provide a comprehensive picture of disease dynamics in companion animals.

Effects of silybin supplementation on nutrient digestibility, hematological parameters, liver function indices, and liver-specific mi-RNA concentration in dogs.

BACKGROUND: Hepatopathies are an important group of disorders in dogs where proper nutritional care is crucial. Supplementation with a hepatoprotectant like silybin can improve liver function and should not interfere with nutrient digestibility. The purpose of this study was to investigate the effect of both pure silybin and commercial hepatoprotectant on nutrients digestibility, liver function indices and health status in healthy dogs (EXP1). Moreover, the second experiment (EXP2) investigated the effect of commercial hepatoprotectant on liver function tests and liver-associated miRNAs concentration in dogs with idiopathic liver disorder. RESULTS: Nutrient digestibility was not affected by treatment in EXP1. Supplementation did alter the serum fatty acid profile, with no clinical relevance. The levels of liver markers such as ALT, AST and GGT significantly decreased. In EXP2, supplementation with commercial hepatoprotectant containing silybin improved liver function tests. A decrease was observed in liver serum markers such as ALT, AST and miR122 concentration. CONCLUSIONS: EXP1 confirmed that silybin (whether pure or as a commercial hepatoprotectant) does not interfere with digestion which subsequently exerts no detrimental effect on dogs' health and metabolism. In EXP2, dietary supplementation with commercial hepatoprotectant containing silybin resulted in a decreased activity of serum liver markers, accompanied by a decrease in the concentration of liver-specific miRNA molecules. Liver function indices were consequently improved. Silybin supplementation can thus serve as an effective therapeutical tool in dogs with hepatopathies.

Screening for structural variants of four candidate genes in dogs with disorders of sex development revealed the first case of a large deletion in NR5A1.

Disorders of sex development (DSD) are important causes of infertility and sterility, and are risk factors for gonadal carcinogenesis. Many DSDs are caused by genetic factors, mainly sex chromosome abnormalities or mutations of genes involved in sexual development, as well as structural variants (SVs) - large deletions, duplications, and insertions, if these overlap genes involved in sex development. The aim of this study was to determine if there were SVs in four candidate genes - NR0B1 (DAX1), NR5A1, RSPO1, and SOX3 - using droplet digital PCR (ddPCR). There was study of two cohorts of dogs with DSD, including 55 animals with XX DSD and 15 with XY DSD. In addition, 40 control females and 10 control males were included in the study. Among cases, for which there were evaluations, a large deletion consisting of four exons of the NR5A1 gene was identified in a Yorkshire Terrier with a rudimentary penis, hypospadias, bilateral cryptorchidism, and spermatogenesis inactive testes. This is the first mutation in the NR5A1 gene leading to XY DSD phenotype to be reported in domestic animals. There were no SVs in the genes evaluated in the present study in the cohort of dogs with XX DSD. The results from this study provide evidence that the large structural variants of these genes are rarely associated with the DSD phenotype in dogs.

Congenital Malformations in a Holstein-Fresian Calf with a Unique Mosaic Karyotype: A Case Report.

A Holstein-Fresian calf with multiple congenital malformations was subjected postmortem to anatomical and genetic investigation. The calf was small (20 kg), had shortened limbs and was unable to stand up. It lived only 44 days. Detailed anatomical investigation revealed the following features: head asymmetry, the relocation of the frontal sinus and eye orbits, hypoplastic thymus without neck part, ductus Botalli, unfinished obliteration in umbilical arteries, and a bilateral series of tooth germs in the temporal region. Cytogenetic examination, performed on in vitro cultured fibroblasts, showed a unique mosaic karyotype with a marker chromosome-60,XX[9 2%]/60,XX,+mar[8%], which was for the first time described in cattle. No other chromosome abnormalities indicating chromosome instabilities, like chromatid breaks or gaps were identified, thus teratogenic agent exposure during pregnancy was excluded. The marker chromosome (mar) was small and it was not possible to identify its origin, however, sequential DAPI/C (4',6-diamidino-2-phenylindole) band staining revealed a large block of constitutive heterochromatin, which is characteristic for centromeric regions of bovine autosomes. We suppose that the identified marker chromosome was a result of somatic deletion in an autosome and its presence could be responsible for the observed developmental malformations. In spite of the topographic distance among the affected organs, we expected a relationship between anatomical abnormalities. To the of our best knowledge, this is the first case of a mosaic karyotype with a cell line carrying a small marker chromosome described in a malformed calf.

Analysis of transcript and methylation levels of INSL3 and RXFP2 in undescended and descended dog testes suggested promising biomarkers associated with cryptorchidism.

Cryptorchidism is the most common disorder of sex development (DSD) in dogs. This malformation is associated with reduced fertility and with a higher risk of gonadal cancer. Testicular descent is a complex process, and the functions of many environmental and genetic factors are crucial for the proper migration of fetal gonads into the scrotum. Among these, the hormone INSL3 (insulin-like peptide 3) and its receptor RXFP2 (relaxin family peptide receptor 2) play crucial roles in the transabdominal migration of the testes. The genetic background of canine cryptorchidism is poorly elucidated. The aim of this study was to compare the transcript and methylation levels of INSL3 and RXFP2 genes in undescended and descended testes of isolated unilateral cryptorchids, and in gonads of control male dogs with scrotal testes. Next, we searched for polymorphic variants in the 5'-regulatory regions of both genes associated with predispositions to cryptorchidism. The INSL3 transcript level was significantly higher in the undescended testes than in the descended testes of both the affected and control dogs. On the other hand, the mRNA level of RXFP2 was significantly lower in the retained gonads of cryptorchids than in the scrotal testes. The methylation level of a single CpG site located 15 bp upstream of the translation start codon in INSL3 was significantly higher in the testes of the control dogs than in both gonads of cryptorchids. The methylation level of 14 CpG sites in the coding region of INSL3 was significantly higher in undescended testes than in the scrotal testes, which may be associated with the higher mRNA levels of INSL3 observed in these samples. The methylation pattern of two CpG sites in the 5'-flanking region of RXFP2 was similar in both descended and undescended testes. We detected three and seven single nucleotide polymorphisms (SNPs) in the 5'-regulatory regions of INSL3 and RXFP2, respectively. Among these, the frequency of A > C substitution (ss7093349755) located 495 bp upstream of the transcription start site of RXFP2 differed significantly between cryptorchids and control dogs. Our study showed two possible genetic biomarkers associated with canine cryptorchidism: a hypomethylation of a single CpG site in the 5'-flanking region of INSL3, and the ss7093349755 SNP in the 5'-flanking region of RXFP2.

Kinetics of single and dual simultaneous infection of pigs with swine influenza A virus and porcine reproductive and respiratory syndrome virus.

BACKGROUND: Simultaneous viral infections exhibit the phenomenon of viral interference, but understanding of the effect of one virus on another is limited. OBJECTIVE: Evaluate and compare clinical characteristics, immune and acute phase response, viral shedding and viral load in pigs singly and doubly inoculated with swine influenza A virus (swIAV) and porcine reproductive and respiratory syndrome virus (PRRSV). ANIMALS: Fifty-four 7-week-old piglets. METHODS: Clinical status and gross lung lesions were scored. Titration of swIAV was carried out in Madin-Darby canine kidney cells. The PRRSV RNA was quantified using a commercial qPCR kit. Antibodies were detected by hemagglutination inhibition assay and commercial ELISA. A lymphocyte proliferation assay was used to measure antigen-specific T-cell responses. Acute phase proteins were determined using ELISA. RESULTS: No differences were found between mean clinical scores, swIAV and PRRSV shedding, and magnitude of the humoral and T-cell response between single-inoculated and dual-inoculated groups. Concentrations of C-reactive protein and haptoglobin increased in PRRSV-inoculated and coinoculated groups, whereas serum amyloid A concentration was increased in groups inoculated or coinoculated with swIAV. Mean swIAV TCID50 titers in the lungs did not differ significantly between coinoculated and swIAV single-inoculated pigs. A significantly higher mean copy number of PRRSV was found in the lungs of PRRSV only-inoculated pigs at 2 day postinoculation (DPI). From 4 DPI, no significant differences in PRRSV load were identified. CONCLUSIONS AND CLINICAL IMPORTANCE: Coinfection of pigs with swIAV and PRRSV did not potentiate clinical signs, lung lesions, immune response, and replication of the viruses in the respiratory tract.

Dynamics of pro- and anti-inflammatory cytokine changes in serum and assessment of their diagnostic utility during lactation impairment in pigs.

Postpartum dysgalactia syndrome (PDS) in sows is a frequent and important clinical problem in the field. Currently, the diagnosis is based on physical examination performed during first days after the farrowing. The present study aimed at evaluation the dynamics of pro- and anti-inflammatory cytokine (IL-4, IL-6, IL-8, IL-10 and TNF-α) changes in serum of sows during peripartum period (day - 28 to + 28) and assessment of their diagnostic utility during lactation impairment in pigs. The study was done on 139 sows divided into 3 groups: clinically healthy sows, sows with lactation disorders, sows which had experienced difficult parturitions, lameness, etc. In order to measure the level of serum cytokines, the quantitative species-specific ELISA assays were used. The investigation demonstrated a different kinetics of changes of studied cytokines in sows from various groups. IL-6 and TNF-alfa shown high dynamic changes after farrowing in in sows. The levels of IL-8 and IL-10 were relatively stable in healthy sows, while in sows with peripartum disorders usually increased during lactation. However, the detailed examination revealed that investigated cytokines cannot be a useful early diagnostic markers of lactation impairments in sow. They do not allow to detect with high probability which sows are susceptible to lactation disorders before the parturition.

Disorders of sex development in cats with different complements of sex chromosomes.

The genetic background of disorders of sex development (DSDs) in cats is poorly understood, due to a relatively low number of such studies in this species. Here we present three new DSD cases with different complements of sex chromosomes. The first, an Oriental Shorthair cat with a rudimentary penis, abdominal atrophic testicles and lack of uterus appeared to be a freemartin, since leucocyte chimerism XX/XY and a lack of Y-linked genes (SRY and ZFY) were observed in DNA isolated from hair follicles. XXY trisomy was identified in the second case, a tortoiseshell Devon Rex male cat with atrophic scrotal testicles and a normal penis. Finally, a European Shorthair cat with atrophic testicles in a bifid scrotum, rudimentary penis and a lack of uterus had XY complement, including Y chromosome of normal size and morphology. Also presence of eight Y-linked genes, detected by PCR, was confirmed. Due to the low testosterone level in this last patient, we searched for a causative mutation in two candidate genes (HSD3B2 and HSD17B3) involved in the metabolism of this steroid hormone. Altogether, five polymorphic sites in HSD3B2 and two in HSD17B3 were found, but none of them showed associations with DSD phenotype. We thus excluded a possibility that the causative mutation is present in these genes. In conclusion, we confirmed that analysis of the sex chromosome complement is a crucial step in diagnosis of DSDs. However, extensive molecular studies of the genes involved in sex development are needed to elucidate the causes of DSDs in cats with normal complements of sex chromosomes.