Gianfranco Militerno

Evidence for bovine besnoitiosis being endemic in Italy - first in vitro isolation of Besnoitia besnoiti from cattle born in Italy

Until 2009, bovine besnoitiosis had never been considered endemic in Italy and the only report on the disease in this country referred to animals imported from France shortly before. However, recently, an autochthonous outbreak of bovine besnoitiosis was reported in four herds located at the intersection of the borders between Emilia-Romagna, Toscana and Marche (Northern Apennine Mountains), which has led to an increased awareness concerning this disease. The present study describes a further outbreak of bovine besnoitiosis in Italy. The afflicted herd was a dairy herd with no evidence for contact with cattle from regions known to be endemic for bovine besnoitiosis. The farm investigation was initiated after a three-year old Holstein Friesian dairy cow with generalized thickening and lichenification of the skin was diagnosed with bovine besnoitiosis. The clinical diagnosis was confirmed by gross pathology, histopathology, serology and PCR. Bradyzoites released from tissue cysts obtained from the skin of this animal enabled the first in vitro isolation of Besnoitia besnoiti in Italy. This isolate was named Bb-Italy1. Sequencing of a 2118 bp spanning region including the complete internal transcribed spacer 1 and parts of the 18S and the 5.8S rRNA gene from DNA extracted from skin-derived zoites revealed a 99.9% identity to sequences known for other B. besnoiti isolated from cattle in Europe. Two GKO mice which had been inoculated intraperitoneally with bovine skin-derived bradyzoites became ill 7 days post inoculation. Parasitophorous vacuoles with multiplying zoites were observed in the cell culture inoculated with peritoneal fluids of these mice and a B. besnoiti infection in the mice and in the cell culture could be confirmed by real-time PCR. A serological investigation in the afflicted herd using immunoblots and an immunofluorescent antibody test (IFAT) revealed an overall herd seroprevalence of 9.7% (31/321), whereas within the female animals older than 2 years 17.0% (29/171) of the dams were tested positive. With one exception, an imported cow from Germany, all the seropositive animals were born in Italy. In connection with previously described autochthonous cases of bovine besnoitiosis the case described herein suggests that bovine besnoitiosis should be considered endemic in Italy.

Evaluation of CALC-I gene (CALCA) expression in tissues of dogs with signs of the systemic inflammatory response syndrome.

Objective – To perform a qualitative evaluation of procalcitonin gene (CALCA) expression in a tissue-specific manner in dogs with signs of the systemic inflammatory response syndrome (SIRS). Design – Observational study. Setting – University Veterinary Teaching Hospital. Animals – Nine clinical cases and 5 research dogs. Interventions – None. Measurements and Main Results – Fresh tissue samples (thyroid, lung, liver, spleen) from 9 dogs that died with a diagnosis of parvoviral infection or SIRS were collected and immediately stored at –80°C. Diagnosis of parvoviral infection was based on clinical signs, positive fecal antigen test, and confirmed by polymerase chain reaction (PCR). Clinical diagnosis of SIRS was based on the clinical criteria reported in veterinary literature. Necropsy was performed on all subjects in the study. Furthermore, thyroid, lung, liver, spleen were collected from 5 normal research dogs immediately postmortem for testing. The 9 dogs with a clinical diagnosis of SIRS died from either parvovirus (n=5), bacterial sepsis (n=3), or neoplasia (n=1). CALCA was amplified by PCR in the following samples: thyroid (9/9), spleen (6/9), lung (4/9), liver (3/9). Only thyroid expressed CALCA in the 5 normal dogs. Conclusions – In SIRS, extrathyroidal transcription of CALCA was documented. Quantitative analysis (real-time polymerase chain reaction) in a wider population of SIRS and normal dogs will provide essential information about the extent and source of extrathyroidal expression of canine CALCA induced by septic and nonseptic systemic inflammation.OBJECTIVE To perform a qualitative evaluation of procalcitonin gene (CALCA) expression in a tissue-specific manner in dogs with signs of the systemic inflammatory response syndrome (SIRS). DESIGN Observational study. SETTING University Veterinary Teaching Hospital. ANIMALS Nine clinical cases and 5 research dogs. INTERVENTIONS None. MEASUREMENTS AND MAIN RESULTS Fresh tissue samples (thyroid, lung, liver, spleen) from 9 dogs that died with a diagnosis of parvoviral infection or SIRS were collected and immediately stored at -80 °C. Diagnosis of parvoviral infection was based on clinical signs, positive fecal antigen test, and confirmed by polymerase chain reaction (PCR). Clinical diagnosis of SIRS was based on the clinical criteria reported in veterinary literature. Necropsy was performed on all subjects in the study. Furthermore, thyroid, lung, liver, spleen were collected from 5 normal research dogs immediately postmortem for testing. The 9 dogs with a clinical diagnosis of SIRS died from either parvovirus (n=5), bacterial sepsis (n=3), or neoplasia (n=1). CALCA was amplified by PCR in the following samples: thyroid (9/9), spleen (6/9), lung (4/9), liver (3/9). Only thyroid expressed CALCA in the 5 normal dogs. CONCLUSIONS In SIRS, extrathyroidal transcription of CALCA was documented. Quantitative analysis (real-time polymerase chain reaction) in a wider population of SIRS and normal dogs will provide essential information about the extent and source of extrathyroidal expression of canine CALCA induced by septic and nonseptic systemic inflammation.

Different biological behaviour of Waldenstrom macroglobulinemia in two dogs

Waldenström Macroglobulinemia is a low-grade immunosecretory disorder associated with lymphoid tumours, which is rarely reported in veterinary medicine. In this study, we describe two clinical cases of this rare syndrome in dogs, each characterized by a different onset and clinical course. In one case, a hyperacute onset and aggressive behaviour of the neoplasm was observed. Absolute serum viscosity (SV) was retrospectively evaluated in order to explain clinical findings. Rotational viscosimetry showed good precision in measuring SV. Both dogs had SV values higher than a control groups of healthy dogs although only one subject developed hyperviscosity symptoms and complications. At high paraprotein concentrations, a slight reduction of the M-component was associated with a marked decrease in SV. Thus, this work suggests that SV assessment is a relevant tool for managing monoclonal gammopathies, whose usefulness should be further confirmed in larger cohorts of dogs.

Congenital facial deformities, ascites and hepatic fibrosis in Romagnola calves

IN RECENT decades, breeders of Romagnola cattle have experienced an increase in the occurrence of a complex malformation, mainly characterised by facial deformities, an enlarged fluid-filled abdomen and hepatic fibrosis ([Gentile and others 2004][1]). This is known by farmers as ‘paunch calf’.

Retroperitoneal Pulmonary Choristoma in a Newborn Calf

A newborn male Romagnola calf, who had died a few hours after birth, was submitted for postmortem examination. Necropsy revealed a 23 cm × 22 cm × 5 cm, pale pink, lobulated, elastic, partially fluctuant mass that protruded from the dorsal retroperitoneal space into the abdominal cavity, extending from the diaphragm to the left kidney. The mass consisted of mature pulmonary tissue and was consistent with a pulmonary choristoma. The gross and microscopic appearance of this rare tumor-like congenital lesion and the possible pathogenesis are discussed.

Cytological features of mandibular actinomycotic osteomyelitis in a cow

ACTINOMYCOTIC osteomyelitis and periostitis of the mandible or maxilla are diseases of cattle caused by the Gram-positive branching filamentous microorganism Actinomyces bovis . The classical lesion, also called ‘lumpy jaw’, was described for the first time by [Bollinger (1877)][1] and more

Comparison between May-Grünwald-Giemsa and rapid cytological stains in fine-needle aspirates of canine mast cell tumour: Diagnostic and prognostic implications

Mast cell tumours (MCTs) are often diagnosed by cytology based on the identification of purple intracytoplasmic granules with methanolic Romanowsky stains, including May-Grünwald-Giemsa (MGG). In clinical practice, aqueous rapid stains (RS) are commonly used, but mast cell granules may not stain properly. Aim of this prospective study was to investigate the frequency of MCT hypogranularity with RS and its potential implications in tumour identification, cytological grading assessment and recognition of nodal metastatic disease. Cytological preparations of canine primary MCTs and metastatic lymph nodes with subsequent histopathological confirmation were included. For each case, good-quality smears were stained with both MGG and RS and comparatively assessed. Eleven of 60 (18.3%) primary MCTs were hypogranular with RS; 9 of them were histologically high-grade tumours and in 3 cases (5%) a definitive MCT diagnosis could not be made. Accuracy in cytological grading assessment (85%) did not differ between RS and MGG. Thirteen of 28 (46.4%) metastatic lymph nodes were hypogranular with RS and 3 independent observers failed to identify nodal MCT metastases in 7% to 18% of RS-stained smears. This study confirms that, in limited cases, RS can be ineffective in staining MCT granules, particularly in high-grade tumours, thus making diagnosis more dependent on experience and quality of preparations. In dubious cases, methanolic stains should be applied. The use of RS is discouraged for the search of nodal metastases, as the identification of isolated mast cells can be more challenging.

Enrichment tools for undocked heavy pigs: effects on body and gastric lesions and carcase and meat quality parameters

Abstract Two independent trials were carried out to assess the effect of two enrichment tools on the prevalence of skin and tail lesions recorded prior of slaughtering, lesions of the pars oesophagea (OGL) of the stomach, and on carcase and meat quality traits in Italian heavy pigs (body weight range: 25–160 kg). Eighty undocked barrows (Landrace × Large White) were used in two trials (20 pigs/experimental group, 5 pigs/pen). In Trial 1, the control group received a hanging metal chain (C1), while the other group received wood logs (WL) placed inside a metal rack. In Trial 2, the control group was provided with hanging chain (C2), while the pen of the other group was enriched with a vegetal edible block (EB) placed inside the metal rack. In both trials, no differences were observed in the prevalence and severity of skin, tail and gastric lesions (p > .05). In Trial 1, WL pigs presented lower backfat (p = .01), higher lean meat percentage (p = .03) and higher drip loss in the loin muscle (p = .02) than C1 pigs. Tail score and gastric lesions showed a moderate correlation (r = 0.42; p = .01) in Trial 1. Treatments had no effect on carcase or meat quality traits in Trial 2 (p > .05). In conclusion, the two enrichments provided did not affect body and gastric lesions, carcase and meat quality of Italian heavy pigs, if compared to the metal chains.

Mislabelling of Döner kebab sold in Italy

The aim of this study was to assess whether Döner kebabs sold in Italy comply with legal labelling requirements. A checklist of EU labelling requirements was drawn up and nine labels of Döner kebab marketed in the province of Modena were collected to evaluate the mandatory information laid down in article 9 of Regulation (EU) No 1169/2011 together with the mandatory requirements of Regulation (EC) 853/2004 for meat preparations. The study showed that different additives are commonly used in Döner kebabs, including E 621 (monosodium glutamate). This additive is not included among the ingredients authorized for the meat preparation food category. Eight frozen Döner kebab samples were taken and a visual assessment and a histologic examination were performed to verify potential changes in the meat’s inner muscle fibre structure that could allow the product to be considered among meat preparations or meat products. The Döner kebab labels showed several systematically deficient regulatory requirements in both extrinsic defects (not related to food composition) and aspects concerning the protection of consumers’ health. Histological examination showed that the tissue was moderately swollen, but the muscle fibre structure was intact. Kebab can be reasonably classified as a meat preparation thereby precluding the use monosodium glutamate (E 621) as an ingredient.

Bilateral Trochlear Nerve Palsy as a Consequence of Cerebellar Medulloblastoma: Clinical and Pathological Findings in a Calf

1-month old female Holstein calf had not been able to stand since birth. Delivery was uncomplicated and no treatment had been administered before admission. On physical examination, the calf had slightly decreased body condition, but was bright and alert. The calf was in lateral recumbency with spastic extension of all four limbs and opisthotonus. If positioned in left lateral recumbency, the calf raised its head, which fell immediately to the ground after rotation of the neck. The calf made no attempt to raise its head when lying in right lateral recumbency. Attempts to passively position the calf in a quadrupedal stance or in sternal recumbency were unsuccessful. Despite the lateral recumbency, the calf was able to suckle. A thorough neurologic examination was carried out. Mental status and consciousness were considered normal. Because of recumbency, postural reactions could not be tested. The menace response was absent in the right eye and present in the left eye. In both eyes, the medial end of the pupil was rotated dorsally. This dorsomedial rotation of the eyes was not affected by changes in the position of the head. Moreover, incomitant strabismus (i.e., degree of misalignment of the eyes varying with the position of