Quantitative studies on B-mode ultrasound and point shear wave elastography of kidneys in nonazotemic dogs.

Renal diseases in dogs can be diagnosed effectively using B-mode ultrasound. Point shear wave elastography (pSWE) has demonstrated usefulness in diagnosing renal diseases in human medicine. However, its application in veterinary medicine is in its nascent stage. It was hypothesized that establishing pSWE reference values in nonazotemic dogs would prove valuable in differentiating renal diseases. In light of this, a single-center, quantitative study with an objective to normalize B-mode ultrasound parameters and pSWE values of the kidney in nonazotemic dogs was conducted. A total of 198 animals presented with clinical signs of anorexia, vomiting, weight loss, and dehydration were enrolled in the study spanning 2 years. Among them, 52 nonazotemic dogs were included as subjects for the study. B-mode ultrasound quantitative parameters, including length (L), breadth (B), height (H), cortical thickness (RCT), and medulla thickness (RMT) of the kidneys, as well as the diameter of the aorta (Ao), were normalized. Additionally, calculated parameters such as L:Ao, B:Ao, H:Ao, RCT:Ao, and corticomedullary ratios were worked out. Point shear wave elastography values were obtained from the cranial and caudal poles of renal cortices using ElastPQ stiffness software. The pSWE values of kidneys in nonazotemic dogs were normalized. The mean ± standard error values were 1.04 ± 0.08 m/s (95% confidence interval: 0.88-1.19 m/s) and 4.18 ± 0.62 kPa (95% confidence interval: 2.93-5.42 kPa). In conclusion, B-mode ultrasound quantitative parameters, ratios, and pSWE values were normalized in nonazotemic dogs, which may prove valuable in differentiating renal pathologies in canine patients.

Rapid detection of Mycobacterium bovis in bovine cytological smears and tissue sections by peptide nucleic acid fluorescence in-situ hybridization.

BACKGROUND: Bovine tuberculosis is the leading cause of death in cattle and other species worldwide. Quick and precise identification of mycobacteria is critical to control the occurrence of tuberculosis in cattle. METHODS: We developed a fluorescent peptide nucleic acid fluorescence in situ hybridization (PNA-FISH) approach to detect Mycobacterium bovis and Mycobacterium avium in cytological smears and tissue sections of bovines suspected of having tuberculosis. PNA-FISH was conducted on smears of lung and lymph node tissues. Standard bovine mycobacterial cultures were used to standardize the probes using 50 % formamide for M. bovis and 30 % formamide for M. avium. M. bovis probe (MTBCcy3), which was standardized at hybridization conditions of (55 °C and 40 % formamide) concentrations, was positive in all cytological smears. RESULTS: Four out of twenty five samples tested positive in tissue sections observed as a bright red fluorescence with a cy3 filter (MTBC probe). No results were observed with (MAVTAMRA) probe for M. avium which was standardized at hybridization conditions of (55 °C and 30 % formamide). No fluorescence was observed in the control tissue sections. Additionally, the results were juxtaposed with those of other commonly used detection methods such as immunohistochemistry and Polymerase Chain Reaction (PCR) by targeting the esxA gene. None of the samples tested positive for M. avium infection. CONCLUSION: PNA-FISH can be used to obtain cytological impression smears and tissue sections. When compared to PCR it consumes less time in the diagnosis of bovine tuberculosis in post mortem cases.

Successful surgical management of massive ovarian teratoma in a rhesus macaque (Macaca mulatta).

A 5 years-old Rhesus macaque was presented with abdominal enlargement. The clinical, radiographic and ultrasonographic findings suggested that a massive mass occupying the whole of the abdomen. The mass was surgically removed, and histopathology confirmed ovarian teratoma. The macaque recovered uneventfully; however, the management to avoid self-mutilation of skin sutures was challenging.

Bioaccumulation of pesticide contaminants in tissue matrices of dogs suffering from malignant canine mammary tumors in Punjab, India.

The unprecedented application of pesticides in Punjab, India during green revolution has lead to an environmental crisis due to the accumulation of persistent organic and pesticide pollutants in the environment and biota of this region. The present study aimed at estimating the abundance of pesticide contaminants in three biological matrices of 36 dogs suffering from malignant canine mammary tumor (mCMT) and 6 tumor free control dogs from Punjab, India. Presence of individual and total pesticides in canine biological samples, age and bodyweight of canine patients was assessed as a potential risk factor for mCMT using logistic regression analysis. Chi-square test was employed to determine tissue-specific accumulations of individual pesticides. Spearman's correlation coefficient was estimated to determine the association between the levels of total pesticides in different tissue matrices and with age and bodyweight of mCMT cases. Gas chromatography-ECD analysis of serum, mammary tissue and adjoining mammary adipose tissue revealed fourteen different pesticides including γ-HCH, α-HCH, dieldrin, aldrin, heptachlor, butachlor, p,p-DDT, o,p-DDT, p,p-DDD, p,p-DDE, L-cyhalothrin, permethrin, fipronil, and fenitrothion. Heptachlor, γ-HCH, aldrin and p,p-DDT were more frequently detected, whereas, p,p-DDE and o,p-DDT were the least common. Differential accumulation of pesticides in tissue matrices, particularly between serum and mammary tissue/adipose tissue was observed. We could not find any association between the total pesticide concentrations among serum, mammary tissue and mammary adipose tissue in mCMT cases. We found that the odds for individual pesticide for serum, mammary tissue and adipose tissue were associated with high uncertainties; however, the total pesticide concentration in mammary tissue was near non-significantly associated with higher risk of mCMT with low uncertainty. Statistically non-significant higher odds of CMT occurrence with increase in age was noticed No association between the concentration of total pesticides in different matrices and age and bodyweight of canine subjects was found.

Circulating level of heat shock protein 27 is elevated in dogs with mammary tumors.

In the current study, we have synthesized canine recombinant Hsp27 in E. coli and raised hyperimmune serum against the protein in mice. Using the hyperimmune serum, an indirect ELISA was developed to estimate circulating levels of Hsp27 in dogs with various types of mammary neoplasia and to compare their levels with those of tumor-free apparently healthy dogs. The developed assay had a high specificity (83.3%) and sensitivity (96.6%) for a cut-off value of 0.45 with respect to histopathological examination in discriminating healthy dogs from those bearing mammary tumors. Serum Hsp27 levels were found to be significantly elevated in tumor subjects (20.33 ± 1.23 ng/ml) as compared to healthy controls (6.56 ± 0.51 ng/ml) and the increase in the Hsp27 levels was irrespective of mammary tumor histotypes. However, dogs with grade-II tumors had higher Hsp27 levels as compared to grade-I types. Therefore, Hsp27 can be exploited as one of the 'neoplastic signatures' of canine mammary tumors.

Immunohistochemical localization of estrogen receptor alpha (ERα) in the oviduct of Indian buffalo during follicular and luteal phases of estrous cycle.

The localization and distribution of estrogen receptor alpha (ERα) in different segments of oviduct of buffalo during follicular and luteal phases of estrous cycle were investigated using immunohistochemistry. Tissue samples from the different segments of oviduct from 12 buffaloes (six each during follicular and luteal phases of estrous cycle) were collected from slaughter house after assessing the gross morphology of ovaries. In addition, blood samples were collected from the animals before slaughter to estimate levels of estrogen and progesterone hormones. The tissue distribution of estrogen receptor was determined by immunohistochemical technique using one-step polymer HRPO staining system. The estrogen receptor was localized in the lamina epithelialis, propria submucosa, tunica muscularis, and tunica serosa. The maximum localization was observed in the lamina epithelialis, where both ciliated and secretory cell types were positive for ERα. Percentage of positive cells varied during the follicular and luteal phases of estrous cycle. The lining epithelium of oviductal glands was also intensely positive for ERα. No immunostaining was observed in any tunic of the oviduct when the primary antibody was replaced by antibody diluent or buffer, and it served as negative control. The data showed that highest immune positive cells were observed in the ampulla region of the oviduct and these cells were lowest in the utero-tubal junction (p < 0.05). Infundibulum, ampulla, and isthmus showed a higher percentage of ERα-positive cells during follicular phase of estrous cycle as compared with those of the luteal phase of estrous cycle (p < 0.05). There was no significant difference in the percentage positive cells during the two phases of estrous cycle in the utero-tubal junction. Immunogold labeling with anti-ERα antibody confirmed the findings of immunohistochemical study at subcellular level. The higher expression during the follicular phase was directly correlated with the level of estrogen hormone.

Over-expression of mammaglobin-B in canine mammary tumors.

BACKGROUND: Mammaglobin, a member of secretoglobin family has been recognized as a breast cancer associated protein. Though the exact function of the protein is not fully known, its expression has been reported to be upregulated in human breast cancer.We focused on studying the expression of mammaglobin-B gene and protein in canine mammary tumor (CMT) tissue. Expression of mammaglobin-B mRNA and protein were assessed by quantitative real-time polymerase chain reaction (qRT-PCR) and immunohistochemistry (IHC), respectively. RESULTS: High levels of mammaglobin-B mRNA expression (6.663 ± 0.841times) was observed in CMT as compared to age and breed matched healthy controls. Further, expression of mammaglobin-B protein was detected in paraffin-embedded mammary tumor tissues from the same subjects by IHC. Mammaglobin-B protein was overexpressed only in 6.67% of healthy mammary glands while, a high level of its expression was scored in 76.7% of the CMT subjects. Moreover, no significant differences in terms of IHC score and qRT-PCR score with respect to CMT histotypes or tumor grades were observed, indicating that mammaglobin-B over-expression occurred irrespective of CMT types or grades. CONCLUSION: Overall, significantly increased expression of mammaglobin-B protein was found in CMTs with respect to healthy mammary glands, which positively correlates to its transcript. These findings suggest that overexpression of mammaglobin-B is associated with tumors of canine mammary glands.

Cerebellar Cortical Abiotrophy in Young Labrador-Retrievers.

Cerebellar abiotrophy is a hereditary degenerative disorder of the central nervous system in humans and animals. Four male and one female Labrador-retriever pups were presented with clinical signs of head swaying with intention tremor, loss of target oriented movement, wide base stance, forelimb hypermetria, and hindlimb ataxia leading to falling off during ambulation. On histopathologic examination of cerebellum, cerebellar cortical abiotrophy was confirmed with extensive loss of Purkinje cells, diminution of granular layer, relative thickening and foliar gliosis in white matter. This is the first reported case of cerebellar abiotrophy in Labrador-retriever to our knowledge.

Molecular and pathological studies on natural cases of bovine theileriosis.

The present communication is a part of study conducted on 32 cases of bovine lymphadenopathies. Out of which, six cases of bovine theileriosis were diagnosed on the basis of peripheral blood smear examination and gross lesions in lymph nodes and visceral organs. Nested PCR using two primer sets N516/N517 and 14136/294 was conducted on samples collected from affected lymph nodes confirms Theileria annulata infection in five out of six cases. Sequencing analysis of amplified product showed 92, 94 and 93 % homology of isolate TH1_Bovine, TH2_Bovine and THEN_Bovine for T. annulata with T. annulata Tamilnadu and Pant Nagar. An upregulation of Th2 cytokines in the cases of theileriosis was observed as the level of TNF-α in individual animals varies from higher value (1028 pg/100 µg protein) to as low as 500 pg/100 µg protein. An increase in level of Th2 cytokines IL-4 and IL-10 was also observed. The present study concluded that pathological studies and cytokine analysis of lymph nodes are of paramount importance in disease diagnosis and associated immune response of the animal with lymphadenopathies.

Histopathological and morphological studies on natural Cysticercus fasciolaris infection in liver of Wistar rats.

Descriptive morphology of multifocal hepatic cysts found in eight of forty five (17.78 %) Wistar rats sacrificed during pharmacological studies related to herbal formulations was studied. The creamish to white cysts were of varying sizes, ranging from 3-8 mm in diameter. Morphological studies of these cysts depicted the presence of metacestodes of Taenia taeniaeformis i.e. Cysticercus fasciolaris inside them. The scolex of metacestode revealed four suckers and rostellum armed with two distinct rows of characteristic pen knife shaped hooks (characteristics of taeniid cestodes). The average size of large hooks was 392.92 ± 10.12 µ and that of small hooks was 240.64 ± 14.26 µ. The average size of suckers was 304.36 ± 12.33 µ. Histopathology of hepatic tissue surrounding the cysts revealed zones of fatty change, inflammation, granulation tissue and metaplasia. However, the histopathology of stomach and small intestines didn't show any significant lesions.

Development of some larval nematodes in experimental and natural animal hosts: an insight into development of pathological lesions vis-a-vis host-parasite interactions.

Infective third-stage larvae of three spiruroid nematodes, Ascarops strongylina and Physocephalus sexalatus of pigs and Spirocerca lupi of dogs, were recovered from 14 species of coprophagous beetles belonging to 4 different genera. These larvae were fed to rabbits and/or guinea pigs to study their development in these experimental hosts. Larvae of A. strongylina reached the adult stage in all rabbits and one guinea pig. The adult worms recovered in these hosts were 40% and 4%, respectively, and became diminutive in comparison to their natural hosts. The larvae of P. sexalatus became reencysted in the gastric wall of rabbits inducing marked pathological changes. The infective larvae of S. lupi became reencapsulated in the stomach wall of the rabbit and also showed development in the aortic wall. Adults of Toxocara canis of dog, collected from 5 different regions of the Indian subcontinent, varied significantly in size. The mouse passage of infective larvae of one of these types led to the recovery of the adults from the experimental dogs that were smaller in size and caused severe pathology in natural experimental hosts. Developmental effects shown in experimental hosts and host specificity are of value in understanding the evolution of nematode parasitism.