Volume 69 (2019): Issue 3 (September 2019)

The aim of this study was to evaluate the main nonconformities that result in cytopathological samples inappropriate for diagnosis in veterinary medicine. All cytopathological samples, obtained from different canine and feline tissues/lesions, included in the study were received and classified as inconclusive by a single public laboratory of veterinary pathology, located in Pernambuco State, Brazil, between 2012 and 2016. Nonconformities regarding the smear quality, cellularity, presence or absence of hemorrhage, cellular overlapping, desiccation, and presence or absence of necrotic debris and/or artifacts were evaluated. Data were tabulated using Microsoft Excel 2007; absolute and relative frequencies were calculated using EPIINFO 3.5.2. From the 3268 cases received between 2012 and 2016, 190 cases were selected and comprised 514 inconclusive slides. The most frequent nonconformities detected were insufficient/absence of cellularity in 100% (514/514), inadequate submacroscopic presentation in 87% (446/514), and hemorrhage in 69% (356/514) of samples. Other features identified were cellular overlapping in 34% (175/514), inadequate staining in 31% (175/514), artifacts in 30% (154/514), desiccation in 28% (145/514), and necrotic debris in 26% (133/514) of samples. The implementation of laboratory standard operational procedures aimed at maintaining quality is essential. It is necessary to initially identify the main errors occurring in the processing stages as a way to guide and design strategies to avoid them.

The primordial germ cells (PGCs) in female animals are comprised of diplotene oocytes arrested in the first meiotic prophase. Expression of Vasa is one of the key factors required for subsequent resumption of development and recruitment of PGCs into the growing follicle class. Since vitamin D regulates recruitment of PGCs and developmental competence of ovarian follicles, this study was designed to investigate the expression of Vasa and rate of apoptosis in foetal ovaries prenatally restricted from dietary vitamin D. Nineteen sexually mature Welsh mountain ewes were randomly assigned to vitamin D deficient (VDD) and vitamin D control (VDC) diets from 17d before mating, up to 125d of gestation, when fetal ovaries were collected and fixed in formalin for immunohistochemistry and TUNEL assay. VDD ovaries had fewer healthy oocytes that could stain positive for Vasa as well as a lower integrated density value for DAB staining intensity. Conversely, TUNNEL staining in VDD animals showed a higher integrated density value and percentage of affected area (P<0.05). The present findings indicate that Vasa expression is decreased, while the rate of apoptosis increased in VDD fetal ovaries, and this may adversely affect resumption of growth and development of PGCs reserve.

Sarcocystosis is a silent, parasitic disease which affects various animal species and causes significant economic losses. It is caused by a number of different intracellular Sarcocystis spp. This study was aimed to detect the host humoral and cellular immune response due to natural infection. Adding to the determination of the infection rate in Monufia Governorate, Egypt. A total number of 127 Egyptian buffaloes (Bubalus bubalis); 30 males and 97 females between 2-11 years of age were examined during 2018. An infection rate of 74% (94/127) was detected by macroscopic examination. The old age females were found to be at a high risk of 90.7% (88/97) in comparison with the young males 20% (6/30). Immunologically, the cellular and humoral immune response was determined using ELISA. A marked down-regulation of the proinflammatory Th-1 cytokine (IFN-γ) and up-regulation of the anti-inflammatory Th-2 cytokine (IL-5) adding to a high level of IgG and IgE were detected in the infected animals compared to the non infected ones. The local cellular immune response in the infected tissues was characterized by an accumulation of mixed inflammatory cells, granuloma formation, eosinophilic infiltration, muscular edema, and necrotic degeneration. In conclusion, the Sarcocystis infection rate in the naturally infected buffaloes in Monufia Governorate was high. This is the first study to provide a fundamental insight into the immune profile in buffaloes infected with Sarcocystis spp. So, it will provide valuable insights to develop novel effective vaccines in future studies. Moreover, sensitive and specific tools should be established for the accurate diagnosis of this disease in the different Egyptian governorates through well-structured serological surveys.

Peripheral nerve sheath tumors (PNSTs) comprise a heterogeneous group of neoplasms originating from the elements of the nerve sheath. They are divided into two forms: benign and malignant PNST. Both benign and malignant PNSTs are not very common in domestic animals but they are reported in different animal species. Histologically, PNSTs are composed predominantly of spindle cells arranged in bundles, whorls and sheets, with a different number of pleomorphic cells and mitotic figures.

The aim of this study was a reclassification of 17 dog tumor samples initially diagnosed with peripheral nerve sheath tumors using histopathological analysis. The main criterion for reclassification was immunohistochemical positivity for various antigens.

PNSTs are often histologically very similar to other spindle cell tumors and immunohistochemistry is required for differential diagnosis. PNSTs generally express vimentin, S-100 protein, glial fibrillary acidic protein (GFAP), collagen IV and laminin.

Four tumor samples were positive to muscular marker α-SMA and vimentin and negative for S-100 protein and desmin. The spindle cells whirling around some blood vessels were observed in these tumors so they were reclassified as perivascular wall tumors (PWTs). The other 13 tumors were S-100 protein and vimentin positive and α-SMA and desmin negative, thus classified as PNST.

The use of the immunohistochemical panel is necessary for distinguishing PNSTs from PWTs in routine diagnostics.

The aim of this study was to clinically and radiologically evaluate the technique of biceps femoris muscle transposition as a new extracapsular treatment technique for cranial cruciate ligament ruptures, which are often encountered in cats. In this study, eight cats diagnosed with cranial cruciate ligament rupture were treated with the biceps femoris muscle transposition technique. The postoperative standard clinical examination procedures were applied to each cat for 90 days. In the preoperative clinical and radiological examinations of the eight cats in the study, cranial cruciate ligament rupture alone was diagnosed in seven of them. Both, the cranial cruciate ligament rupture and meniscal lesions in the same stifle joint were determined in one cat. The biceps femoris muscle transposition technique operation took approximately 20 mins in each case.

The postoperative radiographs were taken on days 10, 30, 60 and 90. No complications were seen in any case during the postoperative follow-up. The Illinois University Evaluation Scale was used for a more objective evaluation. At 90 days postoperatively, there was no lameness in seven out of eight cats, and mild limping was determined in one of them due to concomitant meniscal lesion.

According to the study results, the biceps femoris muscle transposition technique was found to be extremely useful as an easily applicable technique in the treatment of cranial cruciate ligament rupture in cats.

Human activities such as deforestation, urbanization, and environmental pollution lead to a reduction in the spatial boundary between wild animals, domestic animals and humans. These activities increase the risk for the emergence of pathogens from the sylvatic cycle in the population of domestic animals and humans. Foxes are recognized as potential reservoirs for a number of bacterial pathogens of medical and public health concern. The aim of the present study was to investigate the prevalence and spatial distribution of bacterial tick-borne pathogens from the Anaplasmataceae family, Borrelia burgdorferi sensu lato (s.l.), Rickettsia spp., Coxiella burnetii, Francisella tularensis, Bartonella spp., in the red fox population from Serbia and to discuss the obtained results from the epidemiological point of view. Legally hunted red foxes (Vulpes vulpes) from 14 localities in Serbia were included in the study and spleen samples from 129 animals were tested with conventional PCR assays for the presence of bacterial tick-borne pathogens. DNA of Candidatus Neoehrlichia sp. (FU98), Borrelia burgdorferi sensu stricto, Borrelia lusitaniae, and Borrelia garinii was detected in 6 (4.7%), 1 (0.8%), 2 (1.6%) and 1 (0.8%) animals, respectively. Co-infection by Candidatus Neoehrlichia sp. (FU98) and B. garinii was detected in one animal. All samples were negative for other tested bacterial tick-borne pathogens. The results of the present study indicate the potential role of foxes in natural cycles of Candidatus Neoehrlichia sp. (FU98) and causative agents of Lyme borreliosis in the investigated areas. Further research is required to elucidate the role of foxes in the epidemiology of these and other tick-borne zoonotic pathogens in the Republic of Serbia.

Campylobacter species are one of the leading causes of foodborne disease. Poultry is a major reservoir and source of its transmission to humans. The aim of this study was to estimate the prevalence and antimicrobial resistance of Campylobacter spp. isolated from chicken carcasses, the environment, and processing equipment of a poultry slaughterhouse in Greece, to identify the dominant Campylobacter species and to determine if there are clonal relationships among the isolates. Fifty poultry samples and 25 environmental samples were examined using microbial cultures and PCR. Forty-nine of 50 poultry samples (98%) were found to be positive for Campylobacter spp. The environment of the slaughterhouse was also found to be significantly contaminated with Campylobacter spp. Thirty-seven isolates were found to be susceptible to all antimicrobials tested (56.1%) and 29 isolates showed resistance to at least two of the antimicrobials tested (43.9%). We observed 24 different PFGE-types among the 53 isolates with 14 of them isolated only once, while five PFGE-types were represented by two isolates. The remaining 29 isolates were represented by five PFGE-types each consisting of three to 12 isolates. Regarding the relationship of the PFGE types and corresponding resistance profiles, all strains of each PFGE-type shared the same antimicrobial resistance profile. This study reports evidence for Campylobacter spp. cross-contamination among broiler carcasses in a Greek slaughterhouse.

Neurotensin is a tridecapeptide that functions in prenatal metabolism. It is targeted and downregulated by miR-182 through binding the seed site of miR-182 to the 3′ untranslated region and reduced by a candidate tumor suppressor, testin. Considering that apoptosis is critical in the development of mammalian preattachment embryos, this study investigated the modulation of neurotensin to the apoptosis of goat endometrial epithelial cells and apoptosis-related proteins (P38 MAPK and caspase8). Results showed that Neurotensin resisted the apoptosis of goat endometrial epithelial cells through the caspase8 pathway and activated the phosphorylation of P38 MAPK, which is involved in blastocyst formation. Thus, miR-182 is likely to promote uterus health by targeting Neurotensin and upregulating Testin.

The literature in this field cites various ubiquitous fungal and bacterial microorganisms as etiologic agents in severely stressed captive alligators and crocodiles. This study reports two cases of Alligator mississippiensis with bacterial and fungal disease. Two adult American alligators have been submitted for post-mortem investigations. Necropsy, cytology (MGG), and histopathology investigations (HE, HEA, PAS, Gram, Giemsa, Ziehl Neelsen) were carried out. Pleural and pericardial swabs were subjected to microbiological examination. The main lesions detected involved the lower respiratory system and were characterized by thoracic serosanguineous effusions, pleural and pulmonary nodules (1 – 80 mm), accompanied by edema. Similar nodules observed also in the liver, spleen and myocardium, suggested a systemic disease. Additionally, cutaneous, gingival and gastrointestinal erosions and ulcers were found. Cytoarchitecture findings in the major organs revealed lymphoid depletion, multifocal to coalescing necrotic areas with coccoid aggregates and rod shaped bacteria intermixing fungal structures, boarded by heterogeneous inflammatory infiltrates, composed by epithelioid macrophages, lymphocytes and heterophils. The microbiological examination revealed the presence of Aeromonas hydrophila, A. caviae, Serratia marcescens, Pantoea agglomerans, Proteus vulgaris, haemolitic and non-haemolitic E. coli, Citrobacter freundii, Rhizopus/Absidia from pleural and pericardial cavities, concluding that death occurred following a bacterial and fungal pneumonia, with secondary spreading of microorganisms. Along with the low immune response, severe stress was the main possible cause, as a result of environmental temperature changes during the winter, as well as other husbandry issues.

The present paper reports the case of a 3 years old, female Bearded Dragon (Pogona vitticeps) presenting lethargy, anorexia, weight loss, and anemia and marked leukocytosis at CBC. The majority of leukocytes were lymphocytic/lymphoblastic cells (97%). Immunocytochemical staining of blood smears marked for CD3 (neg) and CD79a (pos) suggested immunophenotype B. The patient died after one month from diagnosis. Histology evidenced lymphoid infiltration in the heart, spleen, liver, kidneys and gut. In addition, in the bone marrow a massive infiltration of lymphoid cells confirmed the diagnosis of leukemia. Immunohistochemistry confirmed the CD79a positivity of a large part of infiltrating lymphoid cells indicating a B cells immunophenotype of the neoplastic population. The presence of lymphocytosis and multiorgan infiltration supported the diagnosis of lymphocytic leukemia. Finally, a revision of the literature has also been made.