Abstract. Porcine enzootic pneumonia (PEN), caused by Mycoplasma hyopneumoniae (Mh), has been described in pigs in all geographic areas. The disease is characterized by high morbidity and low mortality rates in intensive swine production systems. A morphologic and immunohistochemical study was done to determine the cellular populations present in lung parenchyma of infected pigs, with special attention to the bronchusassociated lymphoid tissue (BALT). Polyclonal and monoclonal antibodies were used for the detection of antigens of Mh, T lymphocytes (CD3 ϩ , CD4 ϩ , and CD8 ϩ ), IgG ϩ or IgA ϩ lymphocytes, and cells containing lysozyme, S-100 protein, major histocompatibility complex class II antigen or myeloid-histiocyte antigen. Findings in lung tissues associated with Mh infection were catarrhal bronchointerstitial pneumonia, with infiltration of inflammatory cells in the lamina propria of bronchi and bronchioles and alveolar septa. Hyperplasia of mononuclear cells in the BALT areas was the most significant histologic change. The BALT showed a high morphologic and cellular organization. Macrophages and B lymphocytes were the main cellular components of germinal centers. T lymphocytes were primarily located in perifollicular areas of the BALT, lamina propria and within the airway epithelium, and plasma cells containing IgG or IgA at the periphery of the BALT, in the lamina propria of bronchi and bronchioles, in alveolar septa, and around bronchial submucosal glands. The hyperplastic BALT in PEN cases consisted of macrophages, dendritic cells, T and B lymphocytes, and IgG ϩ and IgA ϩ plasma cells. CD4 ϩ cells predominated over CD8 ϩ cells. Local humoral immunity appears to play an important role in the infection.

A new coronavirus known as SARS‐CoV‐2 emerged in Wuhan in 2019 and spread rapidly to the rest of the world causing the pandemic disease named coronavirus disease of 2019 (COVID‐19). Little information is known about the impact this virus can cause upon domestic and stray animals. The potential impact of SARS‐CoV‐2 has become of great interest in cats due to transmission among domestic cats and the severe phenotypes described recently in a domestic cat. In this context, there is a public health warning that needs to be investigated in relation with the epidemiological role of this virus in stray cats. Consequently, in order to know the impact of the possible transmission chain, blood samples were obtained from 114 stray cats in the city of Zaragoza (Spain) and tested for SARS‐CoV‐2 and other selected pathogens susceptible to immunosuppression including Toxoplasma gondii, Leishmania infantum, feline leukaemia virus (FeLV) and feline immunodeficiency virus (FIV) from January to October 2020. Four cats (3.51%), based on enzyme‐linked immunosorbent assay (ELISA) using the receptor binding domain (RBD) of Spike antigen, were seroreactive to SARS‐CoV‐2. T. gondii, L. infantum, FeLV and FIV seroprevalence was 12.28%, 16.67%, 4.39% and 19.30%, respectively. Among seropositive cats to SARS‐CoV‐2, three cats were also seropositive to other pathogens including antibodies detected against T. gondii and FIV (n = 1); T. gondii (n = 1); and FIV and L. infantum (n = 1). The subjects giving positive for SARS‐CoV‐2 were captured in urban areas of the city in different months: January 2020 (2/4), February 2020 (1/4) and July 2020 (1/4). This study revealed, for the first time, the exposure of stray cats to SARS‐CoV‐2 in Spain and the existence of concomitant infections with other pathogens including T. gondii, L. infantum and FIV, suggesting that immunosuppressed animals might be especially susceptible to SARS‐CoV‐2 infection.

Animal infections with SARS-CoV-2 have been reported in different countries and several animal species have been proven to be susceptible to infection with SARS-CoV-2 both naturally and by experimental infection. Moreover, infections under natural conditions in more than 20 mink farms have been reported where humans could have been the source of infection for minks. However, little information is available about the susceptibility of pet animals under natural conditions and currently there is no SARS-CoV-2 epidemiological assessment occurrence in household ferrets. In this study, the presence of SARS-CoV-2 antibodies was evaluated in serum samples obtained from 127 household ferrets (Mustela putorius furo) in the Province of Valencia (Spain). Two ferrets tested positive to SARS-CoV-2 (1.57%) by in-house enzyme-linked immunosorbent assay based on receptor binding domain (RBD) of Spike antigen. Furthermore, anti-RBD SARS-CoV-2 antibodies persisted at detectable levels in a seropositive SARS-CoV-2 domestic ferret beyond 129 days since the first time antibodies were detected. This study reports for the first time the evidence of household pet ferrets exposure to SARS-CoV-2 in Spain to date.

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SUMMARYTwo groups of 32 laying hens (Hyssex Brown) and two groups of 32 23-day-old (Hybro) broiler chickens were fed 2.5 and 5 parts/10 6 of aflatoxin in their diet for 4, 8, 16 and 32 days; 16 hens and 32 chicks were maintained as control groups (0 parts/10 6 ). After the intoxication period, a clearance period was established of 1, 2, 4 and 8 days. Relative weights of liver and kidneys significantly increased in intoxicated hens, but not in broiler chickens. Histological lesions in both types of bird consisted of hepatic cell vacuolation with fatty infiltration. There was a significant decrease (P< 0.001) in egg production in the 5 parts/10 6 group, which started to recover during the clearance period. No morbidity or mortality due to the aflatoxicosis were observed in either type of bird. In intoxicated laying hens, cholesterol levels were not significantly (P> 0.05) different from control values, but triglyceride levels decreased (P< 0.001) in both intoxicated groups. The effect of aflatoxin on calcium and phosphorus levels was important, because on the 4th day their values decreased significantly. Aspartate aminotransferase (AST) serum levels remained normal, whereas alanino aminotransferase (ALT) activity decreased in both intoxicated groups. The activity of serum lactic dehydrogenase (LDH) and gammaglutamil transferase (GGT) increased significantly. In intoxicated broiler chickens, aflatoxins did not alter (P> 0.05) the biochemical parameters studied, except that the serum calcium concentration was lower in the 5 parts/10 6 group. These data indicated that in intoxicated laving hens, a severe clinical biochemical alteration was produced, and that this together with the hepatic lesions observed in hens and broilers may aid disease diagnosis.

A serological and immunohistochemical study of African swine fever was carried out in wild boar killed in seven municipalities in the north of the province of Córdoba during two hunting seasons (1991-92 and 1992-93), when the area was affected by the disease. Fourteen of 147 wild boar analysed by ELISA and immunoblotting had antibodies to African swine fever virus. The immunohistochemical study revealed that four cases (two seropositive and two seronegative) showed immunoreactivity to the anti-VP73 monoclonal antibody. Two of the VP73+ wild boar had severe generalised haemorrhages consistent with the acute from of the disease, and another had lesions consistent with subacute African swine fever, but none of the remaining 144 animals had gross or microscopic changes suggestive of the disease. These results indicate that wild boar can suffer from African swine fever without showing clinical signs. The disease in wild boar was associated with the disease in domestic pigs. Thus, no African swine fever-positive boar were found either in one municipality with no out-breaks in domestic pigs or in three municipalities with only one outbreak in pigs during the hunting seasons and during the previous year. These results suggest that European wild boar do not play an important role as carriers of the virus of African swine fever.

We describe the main pathologic changes in small ruminants affected by AA amyloidosis, together with the partial sequence of the protein involved. Twenty-one sheep and one goat were selected for presenting macroscopic kidney lesions compatible with systemic amyloidosis. Available tissue samples were studied by histologic, immunopathologic, and ultrastructural means. Renal lesions were characterized grossly by pale cortical surfaces with scattered, miliary, whitish-yellow foci and on cut cortical surfaces by straight, whitish-yellow striations. Gangrenous pneumonia was observed in 16 out of 21 affected sheep (76.2%), although other chronic inflammations were also observed. Amyloid was detected in all grossly affected kidneys using Congo red staining, lesions being most remarkable in glomeruli, affecting 95.5% of animals studied. Congophilic deposits were also observed in intertubular interstitium (68.2%) and medulla (57.1%). All amyloid-affected animals presented proximal convoluted tubule lesions, mostly characterized by an increase in diameter and by hyaline granular degeneration that were responsible for the macroscopic appearance of the kidney. Histologically, amyloid was also seen in blood vessels, spleen, liver, lymph nodes, gastrointestinal tract, and adrenal glands. All amyloid deposits demonstrated greenish-yellow birefringence with polarized light, and the antisera prepared against goat amyloid extracts specifically reacted with birefringent congophilic deposits of both sheep and goats. Ultrastructurally, these deposits were formed by masses of straight, nonbranching fibrils located predominantly in the basement membranes of glomerular capillaries and in the mesangium. Partial sequence of the protein in sheep and goats indicated a high degree of homology with the previously reported sequence of sheep Serum Amyloid A.

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