Fatal Simplexvirus humanalpha1 infection in howler-monkeys (Alouatta sp.) under human care: Clinical, molecular, and pathological findings.

BACKGROUND: Simplexvirus humanalpha1 (HuAHV-1) are common anthropozoonosis reported in marmosets but rare in howler monkeys (Alouatta sp.). METHODS: Necropsy of two brown-howler monkeys (A. caraya) and one red-howler monkey (A. guariba clamitans) from different zoo collections were performed. Fragments of all organs were examined through microscopy. Samples were submitted to IHC for Simplexvirus humanalpha 2 (HuAHV-2) [sin. Herpesvirus simplex type 2] and PCR. RESULTS: Grossly, only the A. guariba showed liver lesions characterized by multifocal, pinpoint white areas corresponding microscopically as random necrotizing herpetic hepatitis and ulcerative glossitis. Both A. caraya showed necrotizing meningoencephalitis with Cowdry A-type body inclusions within neurons and astrocytes. Immunolabeling for HuAHV-1/2 was observed in the tongue, liver, and brain. HuAHV-1 was confirmed in all samples by PCR, Sanger sequencing, and phylogenetic analyses. CONCLUSION: Necrotizing meningoencephalitis was appreciated in 2/3 of animals, and it is associated with neurologic signs. Along with ulcerative glossitis, a hallmark lesion in marmosets, it was present in one animal. Regarding herpetic hepatitis, it is not frequent in monkeys and occurs mainly in immunocompromised animals. HuAHV-1 infection was confirmed corroborating with a human source. This is the second report on captive black-howler monkeys and the first gross, histologic, immunohistochemical, and molecular description of herpetic hepatitis and ulcerative glossitis in red-howler monkeys (A. guariba).

Ophthalmic and immunopathological characterization of systemic infectious diseases in cats.

Ocular involvement in systemic diseases is frequent in cats; however, without concurrent clinical and ophthalmic examinations with gross and/or histologic analysis of the eye, these findings can be underdiagnosed. This article aims to provide gross, histologic, and immunohistochemical characteristics of ocular lesions from cats submitted to necropsy, focusing on those caused by systemic infectious agents. Cats that died due to a systemic infectious disease were selected based on necropsy diagnosis and presence of ocular lesions. Gross, histologic, and immunohistochemical findings were recorded. From April 2018 to September 2019, 849 eyes of 428 cats were evaluated. Histologic abnormalities were seen in 29% of cases, which were classified as inflammatory (41%), neoplastic (32%), degenerative (19%), and metabolic/vascular (8%). Macroscopic changes were present in one-third of eyes with histologic lesions. Of these, 40% were attributed to inflammatory or neoplastic diseases associated with infectious agents. The most important infectious agents causing ocular disease in this study were feline leukemia virus, feline infectious peritonitis virus, and Cryptococcus sp. The most common ocular abnormalities associated with infectious agents were uveitis (anterior, posterior, or panuveitis), optic neuritis, and meningitis of the optic nerve. Ocular lesions secondary to systemic infections in cats are frequent; however, these are not always diagnosed because gross lesions are less common than histologic lesions. Therefore, both gross and histologic evaluation of the eyes of cats is recommended, mainly for cases in which the clinical suspicion or necropsy diagnosis suggests that an infectious agent might be related to the cause of death.

Pathological findings associated with Dipetalonema spp. (Spirurida, Onchocercidae) infection in two species of Neotropical monkeys from Brazil.

Among vector-borne helminths, filarioids of the genus Dipetalonema (Spirurida: Onchocercidae) localize in several tissues and body cavities of several animal species, causing mild to moderate lesions. The pathological findings associated with Dipetalonema spp. infection in Neotropical monkeys from southern Brazil are herein described, along with a fatal case due to filarial polyserositis and entrapment of an intestinal segment. At necropsy, nematodes were observed in abdominal and thoracic cavities, or in the pericardium of 37 (31.3%) out of the 118 individuals examined (i.e., 35 Alouatta guariba clamitans and two Sapajus nigritus). In addition, at histology, 27.0% of positive animals presented microfilarie (inside blood vessels of lung, spleen, liver, and brain) and 8.1% presented adult nematodes in the heart, lung, and liver. In two cases, cross-sections of filarioids were associated with areas of epicardial thickening with intense fibrosis and pyogranulomatous inflammation in the brain, heart, liver, lungs, or spleen. The DNA fragment was amplify using the cox1 gene, sequenced and analyzed to identify the nematode species collected; presence of Wolbachia was assessed in the filarioids using the 16S rRNA gene. At BLAST analysis of the cox1 gene, 10 sequences showed 91.7% nucleotide identity with Dipetalonema gracile, and two with D. gracile (98.5%) and Dipetalonema graciliformis (98.3%). Phylogenetic analyses clustered sequences of the cox1 obtained in this study in two clades corresponding with the host species. Wolbachia sp. endosymbiont was detected in four samples. Data herein reported provide a description of pathological lesions associated with the infection by Dipetalonema spp., suggesting that they may cause disease in Neotropical monkeys. In addition, a better understanding of diversity and biology of Dipetalonema spp. in South America is needed to assess the impact they may cause in native non-human primates from Brazil.

Causes of death in neotropical primates in Rio Grande do Sul State, Southern Brazil.

BACKGROUND: Anthropogenic disturbances are the main threats to nonhuman primates conservation, and infectious diseases may also play a key role in primate population decline. This study aimed to determine the main causes of death in neotropical primates. METHODS: A retrospective study of post-mortem examinations was conducted on 146 neotropical primates between January 2000 and December 2018. RESULTS: Conclusive diagnoses were obtained in 68.5% of the cases, of which 59 corresponded to non-infectious causes and 41 to infectious diseases. Trauma was the main cause of death (54/100), with anthropogenic stressors caused by blunt force trauma injuries (collision with vehicles) and puncture wound injuries associated with interspecific aggression (dog predation) were the most common factors. Other causes of death included bacterial diseases (27%), followed by parasitic diseases (12%), neoplasms (2%), and viral diseases (2%). CONCLUSIONS: Free-ranging primates were mostly affected by non-infectious causes, while captive primates were by infectious conditions.

Eosinophilic meningoencephalitis caused by rat lungworm (Angiostrongylus cantonensis) migration in a white-eared opossum (Didelphis albiventris) with concurrent distemper virus in southern Brazil.

Angiostrongylus cantonensis is a worldwide zoonotic parasite that causes eosinophilic meningoencephalitis in many species of animals including humans. This report describes neuro-angiostrongylosis in a white-eared opossum that showed nervous clinical signs such as circling and depression. At necropsy, no relevant macroscopic lesions were observed. Histologically, eosinophilic meningoencephalitis was associated with multiple sections of nematodes and many intracytoplasmic eosinophilic inclusion bodies within gastric parietal cells. Immunohistochemistry was strongly positive for canine distemper virus in the stomach but there was no immunolabeling in the brain. This study describes a fatal case of eosinophilic meningoencephalitis by A. cantonensis with canine distemper virus concurrent infection in a white-eared opossum in southern Brazil, with histological characterization and molecular confirmation of the parasitism.

Outbreak of toxoplasmosis in a flock of domestic chickens (Gallus gallus domesticus) and guinea fowl (Numida meleagris).

Toxoplasmosis is a disease with a worldwide distribution that affects a wide variety of animal species, though with rare descriptions in chickens. We describe the clinical, epidemiological, pathological, and molecular aspects of a toxoplasmosis outbreak in domestic chickens and guinea fowl in southern Brazil. The flock was composed of 47 domestic chickens and 29 guinea fowl. Of these, 22 birds showed clinical signs of lethargy, anorexia, and neurological signs over a clinical course of 24-72 h, and 15 died. Epidemiological data were obtained through fieldwork performed at the chicken farm and necropsies of six birds. Gross lesions were absent at necropsy, and histopathological findings included inflammatory infiltrate of macrophages, lymphocytes, and plasma cells and necrosis in several tissues associated with intralesional Toxoplasma gondii. Immunohistochemistry for T. gondii was positive. Additionally, restriction fragment length polymorphism (RFLP) analysis with 11 markers (SAG1, SAG2 (5'3'SAG2 and alt. SAG2), SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, Apico, and CS3) and microsatellite (MS) analysis with 15 markers (TUB2, W35, TgMA, B18, B17, M33, IV.1, XI.1, N60, N82, AA, N61, N83, M48, and M102) were performed. PCR-RFLP revealed T. gondii genotype ToxoDB-PCR-RFLP #280, and MS analysis also showed a unique genotype. This is the first description of this genotype in chickens and adds to the evidence suggesting considerable genotypic diversity of T. gondii in Brazil.

Pathological and microbiological characterization of mastitis in dairy cows.

Mastitis may be caused by a wide range of microorganisms able to induce distinct lesions in mammary tissues. This study aims to characterize the gross and microscopic features of mastitis in dairy cows and to correlate them with the pathogens involved. The udders of slaughtered dairy cows were inspected and milk samples from each mammary quarter or fragments of the parenchyma were sent for microbiological analysis, and tissue collected for histopathological evaluation. A total of 148 cows and 592 mammary quarters were collected. From these, 432 quarters (73%) had mastitis and in 160 (27%), no changes were observed. Mastitis was classified into seven patterns based on the histopathological findings, of which mixed, lymphoplasmacytic, and suppurative mastitides were the most prevalent with 35.9% (155/432), 27.1% (117/432), and 14.3% (62/432) of the cases, respectively. These patterns were associated with the same set of pathogens: Streptococcus spp., coagulase-negative Staphylococcus (CNS), Staphylococcus aureus, Streptococcus agalactiae, Streptococcus uberis, and Corynebacterium bovis. The pyogranulomatous pattern represented 7.2% (31/432) of the cases with distinct distribution based on the agent involved, mostly S. aureus and Nocardia sp. Abscedative mastitis accounted for 6.0% (26/432) of the cases; it was characterized by multiple abscesses in the parenchyma and was mainly caused by Trueperella pyogenes. Necrosuppurative mastitis represented 5.8% (25/432) of the cases which were characterized by severe parenchyma necrosis and were caused by bacteria such as CNS and Escherichia coli. The granulomatous pattern represented 3.7% (16/432) of the cases and was occasionally associated with Mycobacterium sp.

GASTRIC PYTHIOSIS IN A BACTRIAN CAMEL ( BACTRIANUS CAMELUS).

Pythium insidiosum, an aquatic oomycete, causes chronic lesions in the skin and digestive tract of multiple species. A captive-bred Bactrian camel ( Camelus bactrianus) showed clinical signs of lethargy and weight loss in a clinical course of 30 days, with no response to treatment. At necropsy, the abdominal cavity had approximately 32 L of a yellow, turbid fluid with fibrin. The third compartment of the stomach (C-3) showed a focal area of rupture covered with fibrin. Close to this area, the C-3 wall was thickened and firm, demonstrating irregular, yellow, and friable areas on cut surface (kunkers). Microscopically, these corresponded to necrosis, characterized by a central amorphous eosinophilic material, surrounded by a pyogranulomatous inflammatory infiltrate and fibrosis. Negatively stained hyphae were observed at the periphery of the necrotic areas, which showed marked immunostaining for P. insidiosum. Pythiosis in camelids may involve the stomach, resulting in peritonitis and death.

Pathological findings and patterns of feline infectious peritonitis in the respiratory tract of cats.

Feline infectious peritonitis (FIP) is an important cause of death in cats. Thoracic manifestations are less common than abdominal manifestations, and FIP-associated respiratory disease is poorly documented. This study aimed to investigate pathological findings in the respiratory tract of cats with FIP and the occurrence and distribution of feline coronavirus antigen in the respiratory tract using immunohistochemistry. A retrospective study was carried out on 112 cats with FIP, of which 66 had inflammatory histological lesions in the respiratory tract (58.9%) and were included in this study. Three major gross patterns were defined: marked fibrin deposition in the thoracic cavity with lung atelectasis; marked fibrin deposition in the thoracic cavity with lung pyogranulomas; and lung pyogranulomas without thoracic effusion. Histological analysis revealed primary lesions in the visceral pleura and lung parenchyma at a similar frequency, with multifocal to diffuse presentations. Marked lesions were commonly observed. Five major histological patterns were defined: pleuritis; pleuritis and vasculitis/perivascular injury in the lung parenchyma; pleuritis and pneumonia; perivascular injury in the parenchyma without pleuritis; and pneumonia without pleuritis. In the pleura and pulmonary parenchyma, FIP virus antigen was detected in perivascular and peribronchial macrophages and in macrophages within bronchial-associated lymphoid tissue and foci of necrosis and inflammation in the pleura and lung parenchyma. Co-infections with retroviruses were detected in 47 cats (71.2%), mainly with feline leukemia virus (62.2%). Although FIP is a systemic disease, some cats developed significant lesions in the thoracic cavity, including involvement of the upper respiratory tract and presenting respiratory signs, without other classic signs of FIP. This work advances our knowledge of FIP in the respiratory system, helping veterinarians to recognize the various presentations of this disease.

Pet Pyometra: Correlating Bacteria Pathogenicity to Endometrial Histological Changes.

Pyometra is a life-threatening infectious disease that frequently affects bitches and queens. Although histopathological patterns of pyometra have been extensively explored, the microbiological aspects, such as bacteria pathogenicity, have not been correlated to microscopy endometrial lesions so far. In this study, these two pathological aspects of pyometra were analysed and correlated. Uterus fragments and intrauterine content samples were collected from pets diagnosed with pyometra (30) and submitted to histopathology analysis and bacterial culture, respectively. The degree of endometrial histopathological lesions in pyometra cases were classified as mild, moderate and severe. Thirty different bacteria isolates were identified from intrauterine content culture. Escherichia coli (E. coli) was pure isolated in 57.7% and highly related to severe endometrial lesions. Immunohistochemistry assay revealed the adhesion and invasion of this bacteria agent to the injured endometrium. Virulence aspects of these E. coli strains were explored, demonstrating biofilm formation ability and a set of virulence genes in most isolates. These results support the adaptive genetic and phenotypic advantages of E. coli for uterus infection, and justify the high frequency of this agent involved in pyometra cases.

Rangelia vitalii in free-living crab-eating foxes (Cerdocyon thous) in Uruguay.

Rangelia vitalii is a protozoan parasite that causes a hemorrhagic and hemolytic disease in dogs known as rangeliosis. Current reports of the disease are concentrated in the southern and southeastern regions of Brazil, as well as in Uruguay, Argentina, and Paraguay, and mainly concern domestic dogs. South American wild canids, such as the crab-eating fox (Cerdocyon thous), the pampas fox (Lycalopex gymnocercus), and the maned wolf (Chrysocyon brachyurus) may also be affected, although existing reports are restricted to Brazil. The present study aimed to detect R. vitalii parasitism in the Uruguayan wild fox population. DNA extracted from the blood and/or spleen samples of road-killed C. thous and L. gymnocercus found in northern Uruguay were subjected to polymerase chain reaction (PCR) to amplify a 551-bp fragment of the Rangelia 18S rRNA gene. A total of 62 wild canids, including 38 C. thous and 24L. gymnocercus, were analyzed. Five crab-eating fox samples (13.2%) were positive for R. vitalii, with 99.5-100% identity between the sequences. All samples from pampas fox tested negative for R. vitalii. When compared with the R. vitalii sequences available in GenBank, a similarity of 98.9-100% was revealed. Molecular analysis results suggest that R. vitalii is circulating in the crab-eating fox population in Uruguay; however, its veterinary relevance for these foxes remains unknown.

Feline giant-cell pleomorphic sarcoma: cytologic, histologic and immunohistochemical characterization.

OBJECTIVES: This study aimed to characterize the cytologic, pathologic and immunohistochemical (IHC) aspects of feline giant-cell sarcoma. METHODS: Biopsy and necropsy reports from the Department of Veterinary Pathology were retrieved, and 13 cases of pleomorphic sarcoma (PS) were selected according to the established epidemiologic, pathologic and IHC criteria. All samples were fixed in 10% formalin, routinely processed for histology, and stained with hematoxylin and eosin. Samples also underwent IHC testing for vimentin, ionized calcium-binding adaptor molecule 1 (Iba-1), desmin, actin and S-100. RESULTS: The mean age of the affected cats was 9.5 years, and females were over-represented. Most neoplasms were observed in the flank, lateral thorax, limbs and interscapular region, and were >2 cm in diameter. Cytology analysis revealed highly cellular preparations with three distinct populations (spindle cells, small round cells and multinucleated giant cells) in a dense eosinophilic stroma. Histologically, PS was composed of a combination of these three populations. IHC labeling for vimentin and Iba-1 was strongly positive for spindle cells and multinucleated giant cells, respectively. Desmin/actin showed variable labeling among the samples. S-100 was negative in all samples. CONCLUSIONS AND RELEVANCE: PS is a neoplasm of mesenchymal origin, also known as malignant fibrous histiocytoma. The predominant subtype in this study that affected the cats was the giant-cell type, characterized by the presence of multinucleated giant cells among spindle-shaped cells. These findings are similar to those described in human patients; thus, a comparison between the neoplasms seen in these species might be useful, and the knowledge of biologic behavior and overall treatment approach for humans could be extrapolated to cats.

Rangelia vitalii molecular and histological quantification in tissues comparing crab-eating foxes (Cerdocyon thous) and domestic dogs.

Rangeliosis is a condition transmitted by the tick Amblyomma aureolatum and caused by the protozoan parasite Rangelia vitalii in canids. In domestic dogs, the disease causes a severe hemolytic disease, while in wild canids the piroplasm is often detected without any clinical abnormality. This study aimed to detect and quantify the number of copies of the R. vitalii Hsp70 gene (indirect parasite burden) in several organs of domestic and South American wild canids (Cerdocyon thous and Lycalopex gymnocercus) to elucidate distinct clinical presentations of rangeliosis in these species. A total of seven domestic dogs that died due to rangeliosis and 38 wild foxes were initially included, with all dogs presenting histological and molecular features of rangeliosis, while eight C. thous were positive at the molecular analysis for R. vitalii. Fragments of 22 organs collected from domestic (n = 7) and wild foxes (n = 8) were employed for histological and molecular quantification using real-time polymerase chain reaction aiming at the Hsp70 gene. Histologically, parasitophorous vacuoles were constantly detected in the dogs, while these were detected only in two C. thous. Parasitic burden was significantly higher in the digestive, cardiorespiratory, endocrine, genitourinary, and skeletal-muscle systems of domestic dogs when compared to wild foxes. In the hematopoietic system of wild canids, some organs, such as the lymph nodes and tonsils, presented significantly lower amounts of R. vitalii, while other organs (spleen, bone marrow, and blood) had results similar to those of domestic dogs. Additionally, the central nervous system of both domestic and wild canids presented a similar quantity of R. vitalii. The etiological agent is possibly maintained through an asexual reproductive process (merogony) in both domestic and wild species. Nonetheless, a limited or short-duration schizogony phase occurs in C. thous, which would designate this species as a possible reservoir host for the agent. Dogs, in contrast, would most likely act as accidental hosts, presenting a severe and more pathogenic schizogony phase, resulting in characteristic clinical and pathological rangeliosis.

Relationship Between Clinicopathological Features and Prognosis in Appendicular Osteosarcoma in Dogs.

Osteosarcoma is the most common primary bone neoplasm in dogs and often involves the appendicular skeleton. We report the clinicopathological and prognostic features of appendicular osteosarcomas diagnosed in 153 dogs from 2008 to 2018. The survival data for 22 dogs that underwent surgery and chemotherapy were statistically compared with the clinical, histopathological and immunohistochemical data for correlation with prognosis. The affected dogs had a mean age of 9.1 years and a mean body weight of 33.4 kg. No sex predilection was detected, although the incidence was slightly higher in females (52%). Large mixed and purebred dogs were most commonly affected. Long bones were affected in 124 cases (92.5%; 124/134). The extremities affected within long bones were the metaphysis of the proximal humerus (29%), distal radius (17.7%), distal femur (15.3%) and proximal tibia (7.2%). Histologically, osteoblastic osteosarcoma was most commonly detected (56.9%), followed by chondroblastic (13.7%), telangiectatic (11.8%), fibroblastic (7.8%), giant cell-rich (5.2%) and poorly differentiated neoplasms (4.6%). Regional lymph nodes were available for analysis in 28 out of 70 excisional biopsies from amputated entire limbs. Metastases to regional lymph nodes were observed in 14.3% (4/28) of these cases. Distant metastases were found in 75% (39/52) of cases in which diagnostic imaging or necropsy was carried out. The lungs were the most common site of metastasis (87.2%; 34/39 cases). Weight, age, sex, affected limb or bone, histological classification, mitotic count or histological grade did not influence survival (P >0.05). Neoplasms located at proximal extremities (n = 7) of affected limbs had a tendency for a poorer prognosis than those at distal extremities (n = 15) (P = 0.06). For these cases, the 1-year survival rate was 14.3% compared with 40% for neoplasms at distal extremities. There was no significant difference in the survival rate when amputation (n = 16) or limb preservation (n = 6) techniques were employed with chemotherapy (P = 0.20). The survival period of dogs that had undergone limb amputation, or limb preservation surgery, and chemotherapy was 73-1,185 days, with a mean and median of 376 and 256 days, respectively. Mild to marked cytoplasmic immunolabelling of osteopontin was found in all cases but the intensity (P = 0.66) and percentage of neoplastic cells labelled (P = 0.49) did not influence survival.

Molecular and pathological characterization of teat papillomatosis in dairy cows in southern Brazil.

Teat papillomatosis is caused by different bovine papillomavirus (BPV) types and is especially important for dairy cows, because it results in severe damage to the health and structure of the mammary gland. This work describes the molecular and pathological aspects of teat papillomatosis in dairy cows in southern Brazil. Samples of teat papillomas were collect from 73 slaughtered dairy cows. Fragments of the lesions were collected in individual pools per animal and subjected to PCR using the FAP primer pair and sequencing of the amplification products. Teats with the remaining lesions were fixed in 10% neutral buffered formalin, routinely processed for histopathology, and stained with hematoxylin and eosin (H&E). Papillomatous lesions were characterized by three macroscopic patterns, namely exophytic (5 [6.9%]), flat (29 [39.7%]), and mixed (39 [53.4%]). Histologically, all samples were identified as squamous papillomas. Partial sequencing of the L1 gene resulted in the detection of 8 classical BPV types (BPVs 4, 6, 7, 8, 9, 10, 11, and 12) in 27 samples, 6 previously reported putative BPV types in 17 samples, and 10 putative new BPV types in 15 samples. Four sequences could not be classified, and 10 were negative in the PCR. There was no correlation between the gross pattern and the BPV type identified, and all the samples were characterized by squamous papillomas under histological examination. However, 24 different BPV types were identified, demonstrating high genetic diversity among BPVs associated with teat papillomatosis in dairy cows in southern Brazil.

Detection and phylogenetic characterization of porcine circovirus 2 from pigs in Mozambique.

Porcine circovirus-associated diseases (PCVADs), caused by porcine circovirus 2 (PCV-2), have a significant economic impact on the swine industry worldwide. In Africa, there is little information, to date, regarding the occurrence of PCV-2, and it has not been reported in Mozambique's swine population. We randomly collected mesenteric lymph nodes ( n = 111) from slaughtered pigs from 9 districts in southern Mozambique. PCV-2 DNA was detected in 54% (62 of 111) of the samples and 78% (23 of 31) of the farms. PCV-2 antigen was detected by immunohistochemistry in lymph nodes (6 of 62; 10%) that were positive for PCV-2 by PCR. Histopathologic changes observed in these lymph nodes were lymphoid depletion, multifocal nodal necrosis, and infiltrates of histiocytes and multinucleate giant cells. One positive sample from each district was selected in order to obtain sequences covering the ORF2 region. Five sequences clustered with PCV-2d, of which 3 sequences from Maputo, Namaacha, and Moamba were grouped with PCV-2d-2; 2 sequences from Manhiça and Matola were grouped as PCV-2d-1; and 4 sequences from Boane, Matutuíne, Chibuto, and Xai-Xai were closely related to PCV-2b-1A/B genotypes. Our study indicates that a diversity of PCV-2 viruses is circulating in the Mozambican swine population.

Piglet colibacillosis diagnosis based on multiplex polymerase chain reaction and immunohistochemistry of paraffin-embedded tissues.

Enterotoxigenic Escherichia coli (ETEC) causes diarrhea in pigs, referred to as colibacillosis. The aim of this study was to optimize multiplex polymerase chain reaction (PCR) and immunohistochemistry (IHC) analyses of paraffin-embedded material to detect pathogenic E. coli strains causing colibacillosis in pigs. Multiplex PCR was optimized for fimbriae (F18, F4, F6, F5, and F41) and toxins (types A and B heat-stable toxins [STaP and STb], heat-labile toxin [LT], and type 2 Shiga toxin [STx2e]), and IHC was optimized for an anti-E. coli polyclonal antibody. Samples (132) from pigs received between 2006 and 2014 with clinical and histopathological diagnoses of colibacillosis were analyzed. E. coli was detected by IHC in 78.7%, and at least one virulence factor gene was detected in 71.2%. Pathogenic strains of ETEC with at least one fimbria and one toxin were detected in 40% of the samples in multiplex PCR. The most frequent virulence types were F18-STaP (7.5%), F18-STaP-STb (5.7%), and F4-STaP (3.8%). A statistically significant association was noted between virulence factors F4, F18, STaP, and STb and positive immunostaining results. Colibacillosis diagnosis through multiplex PCR and IHC of paraffin-embedded tissues is a practical approach, as samples can be fixed and stored for long periods before analysis.