Causas de diarrea neonatal y mortalidad de terneros en tambos de Uruguay, caracterización de especies y subtipos zoonóticos de Cryptospordium y estimación de pérdidas económicas nacionales por mortalidad de terneras lecheras

Los objetivos de esta tesis fueron: 1) identificar enfermedades infecciosas/protozoarias de terneras/os lecheras/os asociadas al síndrome de diarrea neonatal, a falla en la transferencia pasiva de inmunidad (FTPI) y a mortalidad en rodeos comerciales, 2) caracterizar especies y subtipos de Cryptosporidium detectados en terneras/os neonatas/os de tambos del país para y determinar su eventual potencial zoonótico y su distancia con fuentes de aguas superficiales colectadas por plantas potabilizadoras de agua para consumo humano y 3) estimar las pérdidas económicas nacionales debidas a mortalidad de terneras lecheras durante el período de crianza para el sector primario en Uruguay. Para el primer objetivo se realizó un estudio de casos y controles de terneros diarreicos (n = 264) y no diarreicos (n = 271) de hasta 30 días de edad de 27 establecimientos. Las heces se analizaron mediante un ELISA de captura de antígeno para Cryptosporidium spp., rotavirus, coronavirus bovino y Escherichia coli F5+, y mediante cultivo selectivo para Salmonella enterica. La sangre/suero se analizó por RT-PCR o ELISA para detección del virus de la diarrea viral bovina. El suero de terneros de ≤ 8 días (n = 95) se evaluó mediante refractometría para determinar la concentración de proteínas totales en suero (PTS). Mediante un cuestionario se registró si los terneros muestreados murieron antes del desleche (período de crianza). Cryptosporidium spp. y rotavirus fueron los únicos patógenos que se asociaron estadísticamente con diarrea. Los terneros diarreicos, los terneros infectados con cualquiera de los patógenos y los terneros infectados con rotavirus tuvieron concentraciones significativamente más bajas de PTS, sugiriendo una FTPI. Los terneros diarreicos tuvieron mayor probabilidad de morir antes del desleche que los no diarreicos. Los terneros diarreicos infectados con S. enterica también tuvieron mayor probabilidad de morir. Para el segundo objetivo se seleccionaron heces de 255 terneros (170 diarreicos y 85 no diarreicos) que habían resultado positivas para Cryptosporidium spp. por ELISA de antígeno, y se procesaron mediante PCRs anidadas dirigidas a los genes 18S rRNA y gp60, seguidas de secuenciación para identificar los subtipos de C. parvum. En 166 muestras se detectaron 7 subtipos de C. parvum de los cuales 5 son zoonóticos. El subtipo IIaA15G2R1 fue el más frecuente (53,6%; 89/166), seguido por IIaA20G1R1 (24,1%; 40/166), IIaA22G1R1 (11,4%; 19/166), IIaA23G1R1 (3,6%; 6/166), IIaA17G2R1 (3%; 5/166), IIaA21G1R1 (2,4%; 4/166) e IIaA16G1R1 (1,8%; 3/166). No hubo diferencias significativas en las proporciones de terneros diarreicos y no diarreicos infectados con ninguno de los subtipos. Se detectaron dos conglomerados espaciales, uno de los cuales se superpuso con Montevideo y la principal planta de potabilización de agua (Aguas Corrientes) que abastece esta ciudad a partir del río Santa Lucía. Los terneros infectados en todos los establecimientos se encontraban dentro de los 20 – 900 m de un curso de agua superficial natural que drenaba los campos, 10 de los cuales fluían hacia seis plantas de potabilización de agua ubicadas entre 9 y 108 km aguas abajo. Cuatro de estos cursos de agua fluían río abajo hacia Aguas Corrientes. Para el tercer objetivo se desarrolló un modelo bioeconómico en Excel para simular la vida productiva de una hembra lechera bajo las condiciones de manejo y alimentación promedio de los establecimientos lecheros uruguayos. El costo total de criar una ternera desde el nacimiento hasta el desleche a los 75 días de vida fue de US$129,74 (US$ 1,77 ternera/día) y desde el nacimiento hasta el primer parto a los 33 meses de vida de US$589,89 (US$ 0,58/día), siendo los costos de alimentación y de mano de obra los principales contribuyentes al costo total durante estos períodos. La edad al repago de 52,7 meses de vida. El lucro cesante (costo de oportunidad) de una ternera que muere durante la etapa de crianza fue, en promedio, de US$2678,6, estimándose una pérdida de US$ 64.382.829 a nivel del sector primario en los años 2014 – 2020. Las diarreas neonatales infecciosas/protozoarias y la FTPI se asocian a morbimortalidad. Los terneros lecheros son reservorios de enteropatógenos zoonóticos y representan una potencial vía de contaminación de cursos de agua superficial captada por plantas potabilizadoras de agua para consumo humano y la mortalidad de terneras en la crianza causa pérdidas económicas multimillonarias para el sector primario en Uruguay

The First Case of Bovine Astrovirus-Associated Encephalitis in the Southern Hemisphere (Uruguay), Uncovers Evidence of Viral Introduction to the Americas From Europe.

Astrovirus species members of the Mamastrovirus genus (family Astroviridae) have been increasingly recognized as neuroinvasive pathogens in various mammals, including humans, mink, cattle, sheep, and pigs. While cases of astrovirus-associated encephalitis have been reported in North America, Europe, and Asia, their presence has never been documented in the Southern hemisphere. This paper describes a case of astrovirus-associated encephalitis in cattle in Uruguay that broadens the geographic distribution and genetic diversity of neuroinvasive astroviruses and provides phylogeographic evidence of viral introduction to the Americas from Europe. A 22-month-old Holstein steer from a farm in Colonia Department, Uruguay developed progressive neurological signs over a 3-days period before dying. Histopathological examination of the brain and proximal cervical spinal cord revealed disseminated, moderate to severe lymphocytic, histiocytic, and plasmacytic poliomeningoencephalomyelitis with neuronal necrosis. A Mamastrovirus strain in the CH13/NeuroS1 clade, that we called bovine astrovirus (BoAstV)-Neuro-Uy, was identified by reverse transcriptase PCR followed by nearly complete genome sequencing. Additionally, BoAstV was detected intralesionally in the brain by chromogenic RNA in situ hybridization within neuronal perikarya, axons and dendrites. Phylogenetic analysis of BoAstV-Neuro-Uy revealed a close relationship to neurotropic BoAstVs within the Virginia/Human-Mink-Ovine clade, which contains a growing cadre of neuroinvasive astroviruses. Analyzing the complete coding region of neuroinvasive BoAstVs sequences available in GenBank, we estimated an evolutionary rate of 4.27 × 10-4 (95% HPD 2.19-6.46 × 10-4) nucleotide substitutions/site/year. Phylogeographic analysis suggests that the common viral ancestor circulated in Europe between 1794-1940, and was introduced in Uruguay between 1849-1967, to later spread to North America and Japan

Systemic granulomatous disease in dairy cattle from Argentina

An outbreak of systemic granulomatous disease of unknown etiology was diagnosed in a dairy herd from Argentina. Eleven out of 211 cows manifested hyperthermia, depression, alopecia, pruritus, decreased milk production and death in most cases. During necropsy, multifocal petechial hemorrhages in glottis and vulva, white multifocal nodules in the liver and splenomegaly with subcapsular petechial hemorrhages were observed. Both kidneys were diffuse pale and enlarged. Systemic granulomatous hepatitis, myocarditis, pancreatitis and nephritis were observed. Water and food samples tested negative for Arsenic and T2 toxin, respectively. Fusarium equiseti was isolated from alfalfa hay samples. Vicia spp. was not consumed by the affected herd and no other cause of vetch-like disease was registered. Other causes of granulomatous lesions (Mycobacterium spp. and fungal infections) were discarded. The systemic granulomatous disease was suggestive of a type IV hypersensitivity reaction. Although the sensitizing agent was not determined, two components of the ration were suspected: cotton seed and bone ash. Both of them were introduced one month prior to the detection of the first affected cow and the disease resolved since they were removed from the diet.EEA BalcarceFil: Odriozola, Ernesto Raul. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; Argentina.Fil: Dorsch, Matías. Universidad Nacional de Mar del Plata. Facultad de Ciencias Agrarias; Argentina.Fil: Caffarena, Rubén Darío. Instituto Nacional de Investigación Agropecuaria (INIA). Plataforma de Investigación en Salud Animal. Universidad de la República. Facultad de Veterinaria; Uruguay.Fil: Moreira, Ana Rita. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; Argentina.Fil: Fernández, Eduardo Luján. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; Argentina.Fil: Morrell, Eleonora Lidia. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; Argentina.Fil: Cantón, Germán José. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; Argentina

Phylogenetic analyses of rotavirus a from cattle in uruguay reveal the circulation of common and uncommon genotypes and suggest interspecies transmission

Uruguay is one of the main exporters of beef and dairy products, and cattle production is one of the main economic sectors in this country. Rotavirus A (RVA) is the main pathogen associated with neonatal calf diarrhea (NCD), a syndrome that leads to significant economic losses to the livestock industry. The aims of this study are to determine the frequency of RVA infections, and to analyze the genetic diversity of RVA strains in calves in Uruguay. A total of 833 samples from dairy and beef calves were analyzed through RT-qPCR and sequencing. RVA was detected in 57.0% of the samples. The frequency of detection was significantly higher in dairy (59.5%) than beef (28.4%) calves (p < 0.001), while it did not differ significantly among calves born in herds that were vaccinated (64.0%) or not vaccinated (66.7%) against NCD. The frequency of RVA detection and the viral load were significantly higher in samples from diarrheic (72.1%, 7.99 log10 genome copies/mL of feces) than non-diarrheic (59.9%, 7.35 log10 genome copies/mL of feces) calves (p < 0.005 and p = 0.007, respectively). The observed G-types (VP7) were G6 (77.6%), G10 (20.7%), and G24 (1.7%), while the P-types were P[5] (28.4%), P[11] (70.7%), and P[33] (0.9%). The G-type and P-type combinations were G6P[11] (40.4%), G6P[5] (38.6%), G10P[11] (19.3%), and the uncommon genotype G24P[33] (1.8%). VP6 and NSP1-5 genotyping were performed to better characterize some strains. The phylogenetic analyses suggested interspecies transmission, including transmission between animals and humans.Fil: Castells, Matías. Universidad de la República; UruguayFil: Caffarena, Rubén Darío. Universidad de la República; UruguayFil: Casaux, María Laura. Universidad de la República; UruguayFil: Schild, Carlos. Universidad de la República; UruguayFil: Miño, Samue. Instituto Nacional de Tecnología Agropecuaria; ArgentinaFil: Castells, Felipe. Universidad de la República; UruguayFil: Castells, Daniel. Universidad de la República; UruguayFil: Victoria, Matías. Universidad de la República; UruguayFil: Riet Correa, Franklin. Universidad de la República; UruguayFil: Giannitti, Federico. Universidad de la República; UruguayFil: Parreño, Gladys Viviana. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación En Ciencias Veterinarias y Agronómicas. Instituto de Virología E Innovaciones Tecnológicas. - Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Pque. Centenario. Instituto de Virología E Innovaciones Tecnológicas; ArgentinaFil: Colina, Rodney. Universidad de la República; Urugua

Complete genome sequence of Campylobacter fetus isolated from a sheep

Campylobacter fetusis an important reproductive pathogen of ruminantsthat occasionally infects humans. Here, we describe the complete circularized genomeof a strain ofCampylobacter fetussubsp.fetusisolated from a sheep. Thefinal assem-bly consisted of a unique contig with a length of 1,849,237 bp

Diagnostic investigation of 100 cases of abortion in sheep in Uruguay: 2015-2021

The aim of this work was to identify causes of abortion through laboratory investigations in sheep flocks in Uruguay. One hundred cases of abortion, comprising 58 fetuses, 36 fetuses with their placentas, and 6 placentas were investigated in 2015-2021. Cases were subjected to gross and microscopic pathologic examinations, and microbiological and serological testing for the identification of causes of abortion, including protozoal, bacterial, and viral pathogens. An etiologic diagnosis was determined in 46 (46%) cases, including 33 (33%) cases caused by infectious pathogens, as determined by the detection of a pathogen along with the identification of fetoplacental lesions attributable to the detected pathogen. Twenty-seven cases (27%) were caused by Toxoplasma gondii, 5 (5%) by Campylobacter fetus subspecies fetus, and 1 (1%) by an unidentified species of Campylobacter. Fourteen cases (14%) had inflammatory and/or necrotizing fetoplacental lesions compatible with an infectious etiology. Although the cause for these lesions was not clearly identified, T. gondii was detected in 4 of these cases, opportunistic bacteria (Bacillus licheniformis, Streptococcus sp.) were isolated in 2 cases, and bovine viral diarrhea virus 1 subtype i (BVDV-1i) was detected in another. Campylobacter jejuni was identified in 1 (1%) severely autolyzed, mummified fetus. BVDV-2b was identified incidentally in one fetus with an etiologic diagnosis of toxoplasmosis. Microscopic agglutination test revealed antibodies against ≥1 Leptospira serovars in 15/63 (23.8%) fetuses; however, Leptospira was not identified by a combination of qPCR, culture, fluorescent antibody testing nor immunohistochemistry. Neospora caninum, Chlamydia abortus, Chlamydia pecorum, Coxiella burnetii and border disease virus were not detected in any of the analyzed cases. Death was attributed to dystocia in 13 (13%) fetuses delivered by 8 sheep, mostly from one highly prolific flock. Congenital malformations including inferior prognathism, a focal hepatic cyst, and enterohepatic agenesis were identified in one fetus each, the latter being the only one considered incompatible with postnatal life. Toxoplasmosis, campylobacteriosis and dystocia were the main identified causes of fetal losses. Despite the relatively low overall success rate in establishing an etiologic diagnosis, a systematic laboratory workup in cases of abortion is of value to identify their causes and enables zoonotic pathogens surveillance.INIA: PL_27 N-23398ANII: FCE_3_2018_1_148540ANII: FSA_1_2018_1_15268

Bovine coronavirus in Uruguay: genetic diversity, risk factors and transboundary introductions from neighboring countries

Bovine coronavirus (BCoV) is a recognized cause of severe neonatal calf diarrhea, with a negative impact on animal welfare, leading to economic losses to the livestock industry. Cattle production is one of the most important economic sectors in Uruguay. The aim of this study was to determine the frequency of BCoV infections and their genetic diversity in Uruguayan calves and to describe the evolutionary history of the virus in South America. The overall detection rate of BCoV in Uruguay was 7.8% (64/824): 7.7% (60/782) in dairy cattle and 9.5% (4/42) in beef cattle. The detection rate of BCoV in samples from deceased and live calves was 10.0% (6/60) and 7.6% (58/763), respectively. Interestingly, there was a lower frequency of BCoV detection in calves born to vaccinated dams (3.3%, 8/240) than in calves born to unvaccinated dams (12.2%, 32/263) (OR: 4.02, 95%CI: 1.81–8.90; p = 0.00026). The frequency of BCoV detection was higher in colder months (11.8%, 44/373) than in warmer months (1.5%, 3/206) (OR: 9.05, 95%CI: 2.77–29.53, p = 0.000013). Uruguayan strains grouped together in two different lineages: one with Argentinean strains and the other with Brazilian strains. Both BCoV lineages were estimated to have entered Uruguay in 2013: one of them from Brazil (95%HPD interval: 2011–2014) and the other from Argentina (95%HPD interval: 2010–2014). The lineages differed by four amino acid changes, and both were divergent from the Mebus reference strain. Surveillance should be maintained to detect possible emerging strains that can clearly diverge at the antigenic level from vaccine strains.Instituto de VirologíaFil: Giannitti, Federico. Instituto Nacional de Investigación Agropecuaria (INIA). Plataforma de Investigación en Salud Animal; Uruguay.Fil: Caffarena, Rubén Darío. Instituto Nacional de Investigación Agropecuaria (INIA). Plataforma de Investigación en Salud Animal. Universidad de la República. Facultad de Veterinaria; Uruguay.Fil: Casaux, María Laura. Instituto Nacional de Investigación Agropecuaria (INIA). Plataforma de Investigación en Salud Animal; Uruguay.Fil: Schild, Carlos. Instituto Nacional de Investigación Agropecuaria (INIA). Plataforma de Investigación en Salud Animal; Uruguay.Fil: Castells, Daniel. Secretariado Uruguayo de la Lana. Centro de Investigación y Experimentación Dr. Alejandro Gallinal; Uruguay.Fil: Riet‑Correa, Franklin. Instituto Nacional de Investigación Agropecuaria (INIA). Plataforma de Investigación en Salud Animal; Uruguay.Fil: Victoria, Matías. Universidad de la República. Centro Universitario de Salto. CENUR Litoral Norte. Laboratorio de Virología Molecular; Uruguay.Fil: Parreño, Viviana. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología. Argentina.Fil: Colina, Rodney. Universidad de la República. Centro Universitario de Salto. CENUR Litoral Norte. Laboratorio de Virología Molecular; Uruguay.Fil: Castells_Matías. Universidad de la República. Centro Universitario de Salto. CENUR Litoral Norte. Laboratorio de Virología Molecular; Uruguay

Detection, risk factors and molecular diversity of norovirus GIII in cattle in Uruguay

Uruguay is a leading exporter of bovine meat and dairy products, and cattle production is one of the principal economic backbones in this country. A main clinical problem faced by livestock farmers is neonatal calf diarrhea (NCD); however, causes of NCD have not been extensively studied in Uruguay. Bovine norovirus (BoNoV) has been proposed as one of the possible etiologies of NCD as experimentally infected calves developed diarrhea and enteropathy, although limited information is available from field surveys. The aims of this study were to determine the frequency of infection, to investigate possible risk factors, and to determine the molecular diversity of BoNoV in Uruguay. A total of 761 samples of feces or intestinal contents from dairy and beef calves were analyzed through RT-qPCR. The overall frequency of detection of BoNoV was 66.1% with higher frequency in dairy (70.5%) than beef (15.9%) calves (p < 0.01). BoNoV was detected similarly in diarrheic (78.8%) and non-diarrheic (76.2%) dairy calves (p = 0.50). Calves ≤2 weeks of age (84%) were infected more often than older (62.7%) calves (p < 0.01). Phylogenetic analysis confirmed the presence of GIII.1 and GIII.2 genotypes. In addition, we reported the circulation of recombinant strains and the detection of a strain with the recently described novel VP1 genotype. This study represents the first report describing the circulation, the associated risk factors, and the molecular diversity of BoNoV in Uruguay.Instituto de VirologíaFil: Castells, Matías. Universidad de la República. Centro Universitario de Salto. CENUR Litoral Norte. Laboratorio de Virología Molecular; Uruguay.Fil: Castells, Matías. Instituto Nacional de Investigación Agropecuaria (INIA). Estación Experimental La Estanzuela; UruguayFil: Caffarena, Rubén Darío. Instituto Nacional de Investigación Agropecuaria (INIA). Estación Experimental La Estanzuela; UruguayFil: Caffarena, Rubén Darío. Universidad de la República. Facultad de Veterinaria; Uruguay.Fil: Casaux, María Laura. Instituto Nacional de Investigación Agropecuaria (INIA). Estación Experimental La Estanzuela; UruguayFil: Schild, Carlos. Instituto Nacional de Investigación Agropecuaria (INIA). Estación Experimental La Estanzuela; UruguayFil: Castells, Felipe. Profesional independiente; UruguayFil: Castells, Daniel. Secretariado Uruguayo de la Lana. Centro de Investigación y Experimentación Dr. Alejandro Gallinal; Uruguay.Fil: Victoria, Matías. Universidad de la República. Centro Universitario de Salto. CENUR Litoral Norte. Laboratorio de Virología Molecular; Uruguay.Fil: Riet‑Correa, Franklin. Instituto Nacional de Investigación Agropecuaria (INIA). Estación Experimental La Estanzuela; UruguayFil: Giannitti, Federico. Instituto Nacional de Investigación Agropecuaria (INIA). Estación Experimental La Estanzuela; UruguayFil: Parreño, Viviana. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina.Fil: Colina, Rodney. Universidad de la República. Centro Universitario de Salto. CENUR Litoral Norte. Laboratorio de Virología Molecular; Uruguay