Mycobacterium avium subsp. paratuberculosis (MAP) molecular diversity in cattle, sheep, and goats from Latin America and the Caribbean: a systematic review.

This study aimed to systematically collect and appraise the scientific evidence to answer the research question: What MAP genotypes have been isolated from cattle, sheep, and goats in Latin America and the Caribbean? An electronic search was conducted on three platforms (i.e., OVID®, Web of Science®, SciELO) as well as on the proceedings of the International Colloquium on Paratuberculosis. Inclusion and exclusion criteria were defined a priori and conserved through the systematic process and only articles published in peer-reviewed journals were considered. A total of 26 articles met the definitive inclusion criteria. All were published in English, in 15 different journals, and between 1989 and 2020. The relevant articles reported the use of six different genotyping techniques (i.e., polymerase chain reaction-restriction endonuclease analysis, restriction fragment length polymorphism, type-specific-PCR, mycobacterial interspersed repetitive units-variable number of tandem repeats, multi-locus short sequence repeat, single nucleotide polymorphism) in isolates from seven countries. Genotypes found so far in the region using typing techniques were mainly C type. MIRU-VNTR mostly reported INMV 1, INMV 2, and INMV 11 subtypes, among others. MLSSR reported genotypes from four different countries, reporting nine different subtypes of which 7g-10g-4ggt was the most common for loci 1, 2, and 8, respectively. Regardless the high diversity of techniques used so far to genotype Latin American and Caribbean MAP isolates, the original question of this systematic review has been answered. In addition, a relative genetic similarity between MAP strains recovered from cattle, goats, and sheep unrelatedly of the matrix and geographic origin was identified.

Control of paratuberculosis: who, why and how. A review of 48 countries.

Paratuberculosis, a chronic disease affecting ruminant livestock, is caused by Mycobacterium avium subsp. paratuberculosis (MAP). It has direct and indirect economic costs, impacts animal welfare and arouses public health concerns. In a survey of 48 countries we found paratuberculosis to be very common in livestock. In about half the countries more than 20% of herds and flocks were infected with MAP. Most countries had large ruminant populations (millions), several types of farmed ruminants, multiple husbandry systems and tens of thousands of individual farms, creating challenges for disease control. In addition, numerous species of free-living wildlife were infected. Paratuberculosis was notifiable in most countries, but formal control programs were present in only 22 countries. Generally, these were the more highly developed countries with advanced veterinary services. Of the countries without a formal control program for paratuberculosis, 76% were in South and Central America, Asia and Africa while 20% were in Europe. Control programs were justified most commonly on animal health grounds, but protecting market access and public health were other factors. Prevalence reduction was the major objective in most countries, but Norway and Sweden aimed to eradicate the disease, so surveillance and response were their major objectives. Government funding was involved in about two thirds of countries, but operations tended to be funded by farmers and their organizations and not by government alone. The majority of countries (60%) had voluntary control programs. Generally, programs were supported by incentives for joining, financial compensation and/or penalties for non-participation. Performance indicators, structure, leadership, practices and tools used in control programs are also presented. Securing funding for long-term control activities was a widespread problem. Control programs were reported to be successful in 16 (73%) of the 22 countries. Recommendations are made for future control programs, including a primary goal of establishing an international code for paratuberculosis, leading to universal acknowledgment of the principles and methods of control in relation to endemic and transboundary disease. An holistic approach across all ruminant livestock industries and long-term commitment is required for control of paratuberculosis.

Milk yield and lactation stage are associated with positive results to ELISA for Mycobacterium avium subsp. paratuberculosis in dairy cows from Northern Antioquia, Colombia: a preliminary study.

Paratuberculosis is a slow-developing infectious disease characterized by chronic granulomatous enterocolitis. This disease has a variable incubation period from 6 months to over 15 years and is caused by Mycobacterium avium subsp. paratuberculosis (MAP). Some studies have been conducted in cattle during the last decades in Colombia. However, those studies were designed using relatively small populations and were not aimed to establish prevalence. This study aimed to determine the MAP seroprevalence in selected dairy herds and to explore risk factors associated with the serology results. Serum samples and related data were collected from 696 randomly selected bovines in 28 dairy herds located in 12 different districts in one of the main dairy municipalities in Colombia (San Pedro de los Milagros). The samples were analyzed using a commercial ELISA kit. The information on risk factors was analyzed using a logistic regression. The apparent seroprevalence was 3.6 % (1/28) at the herd level and 2 % (14/696) at the animal level. The number of days in milk production between 100 and 200 days, and over 200 days as well as the daily milk production between 20 and 40 L/cow, and over 40 L/cow were associated with MAP seropositivity with odds ratios of 4.42, 3.45, 2.53, and 20.38, respectively. This study demonstrates the MAP seroprevalence in dairy herds from Antioquia, Colombia and the possible relationship between MAP seropositivity, milk yield, and lactation stage.

Systematic review of the prevalence of paratuberculosis in cattle, sheep, and goats in Latin America and the Caribbean.

Mycobacterium avium subsp. paratuberculosis causes paratuberculosis or Johne's disease (JD) in domestic ruminants and wild species. The aim of the present study was to systematically review the prevalence of paratuberculosis among farmed animals (cattle, sheep, and goats) in Latin America and the Caribbean. The initial search for existing publications reporting systematic reviews and primary studies was carried out by searching the available databases. For the final selection of studies, an initial screen for basic eligibility and a detailed appraisal of quality were performed. After study selection, the relevant data were extracted. The detailed appraisal generated 24 publications that reported 52 studies, of which 73.1, 11.5, and 15.4 % were from cattle, sheep, and goats, respectively. Thirty-three (63.5 %) of the studies were animal level studies, while 19 (36.5 %) were herd-/flock-level studies. No flock-level studies on prevalence in sheep were found. Studies in Latin American and Caribbean countries revealed an overall prevalence of 16.9 (95 % CI (confidence interval) 13.2-20.5) and 75.8 % (95 % CI 50.1-101.5) in cattle at the animal and herd levels, respectively; the prevalence was 16 % (95 % CI 7.9-24.1) in sheep at the animal level and 4.3 % (95 % CI 1.9-6.8) and 3.7 % (95 % CI 0.1-7.4) in goats at the animal and flock levels, respectively. In general, prevalence results reported by the studies were insufficient to accurately determine the prevalence of paratuberculosis in farmed animals in Latin America and the Caribbean. Several flaws in the design of studies limit the quality of evidence regarding the prevalence of paratuberculosis in the region.

Extended-spectrum beta-lactamase-producing Klebsiella pneumoniae and risk factors associated with high total bacterial count in bulk tank milk from dairy farms in Colombia.

The objective of the study was to evaluate the frequency and genetic characteristics of ESBL-producing Escherichia coli and Klebsiella spp. and the risk factors associated with a high total bacterial count in bulk tank milk samples of dairy farms in three municipalities of the Antioquia Department, Colombia. Fifteen samples were positive for E. coli and Klebsiella spp. Subsequent analysis of the 16 S rRNA gene sequences confirmed these isolates included E. coli (n = 3), K. oxytoca (n = 11), and K. pneumoniae (n = 1). None of the isolates was positive for ESBL identification by phenotypic methods, but the only the isolate of K. pneumoniae was positive for the blaSHV61 gene by sequence analysis. The antibiotic susceptibility evaluation for all Klebsiella spp. isolates identified resistance to fosfomycin (50%; 6/12) and ampicillin (100%; 12/12). While most of the herds maintain adequate hygienic quality, specific risk factors such as having more than 60 milking cows, frequent changes in milkers, milking in paddocks, and using a chlorinated product for pre-dipping have been identified as associated with a high total bacterial count > 100,000 CFU/mL in bulk tank milk. However, certain variables including the milker being the owner of the animals and the proper washing and disinfection of the milking machine contribute to maintain a high level of hygiene and quality in the raw milk stored in the tanks. In conclusion, the frequency of ESBL producers was relatively low, with only K. pneumoniae testing positive for the blaSHV ESBL type. The presence of these bacteria in milk tanks represents a potential risk to public health for consumers of raw milk and its derivatives.

Staphylococcus aureus, Escherichia coli, and Klebsiella spp. prevalence in bulk tank milk of Colombian herds and associated milking practices.

Background and Aim: Bovine mastitis is one of the most costly and prevalent diseases in dairy herds, which can be prevented and controlled through proper milking practices, diagnosis, and elimination of chronic animals, among others. Contagious pathogens such as Staphylococcus aureus and environmental pathogens such as Escherichia coli and Klebsiella spp. can affect cows and milk for human consumption, generating a public health risk. This study aimed to estimate the prevalence of herds with somatic cell count (SCC) ≥200,000 cells/mL, S. aureus, E. coli, and Klebsiella spp., in bulk tank milk (BTM) and its associated risk factors in Colombian dairy cow herds. Materials and Methods: A cross-sectional probabilistic study was carried out in 150 dairy herds located in the north of the Antioquia province. A single visit per herd was conducted, during which three BTM samples were aseptically collected. General data and milking practices were collected through an epidemiological survey applied in each herd. Results: The prevalence of S. aureus, E. coli, and Klebsiella spp. were 14% (21/150), 2% (3/150), and 8% (12/150), respectively. Moreover, 95% of the herds presented an SCC of ≥200,000 cells/mL. Practices such as in-paddock milking, change of milker during the last month, use of disposable gloves, and hand disinfection were associated with increased prevalence of S. aureus, whereas proper dipping was a protective factor. Proper washing and disinfection of the milking machine, use of chlorinated disinfectants for hand hygiene, and use of disposable gloves decreased the prevalence of E. coli and Klebsiella spp. Bulk tank SCC increased in herds with 30-60 milking cows, herds with >60 milking cows, and herds with a change of milker during the last month. Hand disinfection and dipping decreased the SCC. Conclusion: Staphylococcus aureus, E. coli, and Klebsiella spp. were prevalent in BTM from dairy cow herds. The risk of S. aureus isolation was higher in herds with an in-paddock milking system. The risk of E. coli and Klebsiella spp. isolation were higher in herds with >60 milking cows, with a change of milker during the last month. Processes such as avoiding changing the milker and greater control in medium and large herds could improve the SCC in BTM.

Genotypes of Mycobacterium avium subsp. paratuberculosis from South American countries determined by two methods based on genomic repetitive sequences.

Mycobacterium avium subsp. paratuberculosis (MAP) is the causal agent of Johne's disease or paratuberculosis of ruminants and has been associated with Crohn's disease in humans. In this study, the genotypes of MAP obtained so far in South American countries using a combination of the subtyping methods Mycobacterial Interspersed Repeats Units-Variable Number of Tandem Repeats (MIRU-VNTR) and Multilocus Short Sequence Repeats (MLSSR) were analyzed. Through this analysis, seven different MIRU-VNTR genotypes and seven MLSSR genotypes have been detected. If both methods were combined, nine different genotypes were found. Results revealed the predominance of MIRU-VNTR genotype 1 (INMV 1) and MLSSR genotype A (7 g-10 g-4ggt) among MAP isolates from different host species in South America. These predominant MAP genotypes are also commonly detected in Europe and the United States. This predominance could be the result of higher animal infection ability or better culturability on solid media used for isolation. Further studies on molecular epidemiology of MAP must be carried out in South America to increase our knowledge of the global distribution of MAP.

Anisakidae parasites in frozen fish fillets intended for human consumption. / Parásitos anisákidos en filetes de pescado congelado destinados a consumo humano

INTRODUCTION: Anisakiasis is a human parasitic disease caused by the consumption of raw fish or shellfish containing larvae of the Anisakidae family. It is currently considered an emerging disease of public health interest. OBJECTIVE: To identify the presence of larvae of the Anisakidae family in samples of frozen raw fish fillets intended for human consumption in markets in Medellín and its metropolitan area in Antioquia, Colombia. MATERIALS AND METHODS: A cross-sectional study was carried out, in which larvae of the Anisakidae family were detected and identified in frozen raw fish fillets from three representative markets in Medellín and its metropolitan area. A total of 384 ready for consumption fillets were analyzed (197 sawfish, 137 salmon, 37 tuna, and 13 hake), using the pressing and ultraviolet light method. Taxonomic keys were used to identify the collected parasites and to establish its genus. Conventional PCR and Sanger sequencing was performed to determine the species. RESULTS: Four larvae were found in 4 of the 384 (1.04%) fillets (CI95% 1.04 ± 1.01%). The species of fish in which the larvae were found was sawfish (Scomberomorus spp.) and the genus and species of the larvae was established as Anisakis pegreffii. CONCLUSIONS: According to the study, the presence of Anisakis parasites in frozen raw fish fillets in the influence area is evident.

Introducción. La anisakiasis es una infección producida por parásitos de la familia Anisakidae, transmitida a los humanos por el consumo de pescado o mariscos crudos. En la actualidad, se considera una enfermedad emergente de interés en salud pública. Objetivo. Identificar la presencia de larvas de la familia Anisakidae en muestras de filetes de pescado crudo congelado destinados a consumo humano, en mercados de Medellín y su área metropolitana en Antioquia (Colombia). Materiales y métodos. Se realizó un estudio transversal, en el cual se buscó la presencia de larvas de la familia Anisakidae en filetes de pescado crudo congelado de tres mercados representativos de Medellín y su área metropolitana. Se analizaron 384 filetes listos para el consumo (197 sierras, 137 salmones, 37 atunes y 13 merluzas). Cada filete fue analizado mediante el método de prensado y luz ultravioleta. Los parásitos recolectados se identificaron a partir de claves taxonómicas para establecer el género, así como PCR convencional y posterior secuenciación Sanger, para determinar la especie. Resultados. Se encontraron 4 larvas en 4 de los 384 filetes (1,04 %) (IC95% 1,04 ± 1,01 %). Las larvas encontradas fueron identificadas como Anisakis pegreffi y el tipo de pescado en el cual se encontraron fue la sierra (Scomberomorus spp.). Conclusiones. De acuerdo con el estudio realizado, se evidencia la presencia de parásitos anisákidos en filetes de pescado crudo congelado en el área de influencia.

Paratuberculosis control strategies in dairy cattle: A systematic review.

Background: Mycobacterium avium subsp. paratuberculosis is the causative agent of paratuberculosis (PTB), incurable enterocolitis, affecting domestic and wild ruminants. Economic losses, impacts on animal health and welfare, and public health concerns justify its herd-level control. Aim: To systematically collect information to answer: What are the control and eradication strategies of PTB in dairy cattle worldwide? Methods: The search procedure was carried out on October 2nd, 2019, and updated on August 3rd, 2021, using OVID®, SciELO, and Redalyc databases, and the registers from the International Colloquium on Paratuberculosis (1991-2018). The inclusion criteria considered articles published in English, Portuguese, and Spanish and in peer-reviewed journals. The exclusion criteria included irrelevant topics, species other-than bovines, and not original articles. Definitive studies were obtained through the consensus of the authors on eligibility and quality. Data extraction was performed, considering bibliographic information, control and outcome strategies, follow-up time, and results. Results: Twenty-six relevant studies were found, reporting the use of three grouped control strategies: hygiene and management strategy (HMS), test-and-cull strategy (TCS), and vaccination strategy (VS). The HMS was the most common one (20/26), followed by TCS (17/26) and VS (7/26). Combined control strategies such as TCS-HMS (12/26), TCS-VS (1/26), and HMS-VS (1/26) were also described, and the consideration of the three control strategies (TCS-HMS-VS) was reported in two articles. The HMS included practices such as neonates/juvenile livestock hygiene, biosecurity, prevention of infection introduction into the herd, and environmental management. Within HMS, the most frequent practices were to remove calves from their dams as soon as possible after birth and to keep the minimal exposure of calves and heifers to adult cattle. As limitations, within the HMS, it is considered that some strategies cannot be included due to lack of compliance, or the application of the same strategy among one study and another may have a different degree of interpretation; publication bias was not controlled since the results of the control programs in endemic countries may be not available. Conclusion: The main PTB control strategies in dairy cattle worldwide are HMS, TCS, and VS. The use of one or several combined strategies has been found to succeed in controlling the disease at the herd-level.

Serological and molecular detection of Mycobacterium avium subsp. paratuberculosis in cattle of dairy herds in Colombia.

The objective of this study is the detection of Mycobacterium avium subsp. paratuberculosis (MAP) by serum enzyme-linked immunosorbent assay (ELISA), fecal polymerase chain reaction (PCR), and fecal culture in Colombian dairy herds. Serum and fecal samples from asymptomatic cows (n = 307) of 14 dairy herds were tested for MAP by an unabsorbed ELISA test (ELISA-A). Serum and fecal samples from positive ELISA-A animals (n = 31) were further tested by an absorbed ELISA test (ELISA-B) and PCR. Fecal samples from animals of herds positive by ELISA-A and PCR (n = 105) were inoculated onto three different culture media. ELISA-A produced positive results in 10% of the serum samples and 71% of the herds. ELISA-B and PCR results were positive in two and six serum and fecal samples from positive ELISA-A animals, respectively. Fecal samples were negative for MAP on all culture media. The results of this study confirmed the presence of MAP in local dairy herds and the difficulties of MAP detection in asymptomatic animals by ELISA, PCR, and fecal culture.

Ehrlichia canis TRP36 diversity in naturally infected-dogs from an urban area of Colombia.

Ehrlichia canis is the etiologic agent of a highly prevalent tick-borne disease, canine monocytic ehrlichiosis (CME). Four defined E. canis genotypes based on the trp36 gene sequences have been reported, three of them identified in North or South America. The diversity of E. canis has been investigated using genetic and serologic approaches based on distinct 36 kDa tandem repeat protein (trp36) gene sequences that have been reported. The main objectives of this study were to determine the prevalence of E. canis infection in dogs from Medellín, Colombia by PCR and determine the E. canis diversity using molecular and serologic approaches. Blood was collected from dogs (n = 300) with clinical signs of CME for PCR detection of E. canis 16S rRNA, dsb and trp36 DNA. Phylogenetic analysis of trp36 gene sequences was performed using MEGA. A serological evaluation was performed using immunofluorescence microscopy and ELISA with species-specific peptides from E. canis TRP19 and TRP36 (3 genotypes) and E. chaffeensis (TRP32). E. canis DNA (16S rRNA and/or dsb) was detected in 18 % (53/300) of dogs by PCR amplification. The trp36 gene was amplified and sequenced from 35/53 16S rRNA/dsb PCR positive samples revealing three genotypes: United States (US; n = 21), Costa Rica (CR; n = 11), and Brazil (BR; n = 3). Most dogs (33/35) with detectable trp36 DNA had anti-E. canis TRP19 and TRP36 peptide antibodies that corresponded to the genotype detected by PCR. Dogs that had antibodies to the TRP19 peptide (82/300; 38 %), also had antibodies to one or more genotype-specific TRP36 peptides. Based on TRP36 serology, the dogs exhibited highest frequency of infection with the US genogroup (US = 26), followed by the CR genogroup (CR = 19) and the BR genogroup (BR = 11). Notably, 26/53 trp36 PCR positive dogs had detectable antibodies to multiple E. canis genotypes (US/BR/CR = 8, BR/CR = 7, US/CR = 6 and US/BR = 5) suggesting coinfection or multiple sequential infections with different genotypes. Colombian dogs did not have antibodies to E. chaffeensis as determined by a TRP32 species-specific ELISA. Our results demonstrate the presence of three previously defined genotypes in North and South America in Colombian dogs (US, BR, CR). These results also demonstrate that TRP19 and TRP36 serology can provide valuable information regarding E. canis exposure and the potential genotype(s) involved in infection.

Prevalence of Mycobacterium avium subsp. paratuberculosis infection in dairy herds in Northern Antioquia (Colombia) and associated risk factors using environmental sampling.

This cross-sectional study aimed to determine Mycobacterium avium subsp. paratuberculosis (MAP) herd-level prevalence using a quantitative real-time PCR method (qPCR), performed on environmental samples. Secondly, the study aimed to explore herd-level risk factors associated with the presence of MAP in dairy herds with in-paddock milking facilities of the Northern region of the Province of Antioquia (Colombia). Study herds (n = 292) located in 61 different districts from six municipalities were randomly selected amongst 7794 dairies registered in the foot-and-mouth disease vaccination records from 2015. The sampling strategy considered a proportional allocation, both at municipality and district level. Participant herds were visited once between June and October 2016 to collect one composite environmental sample and to complete a risk assessment questionnaire. Each composite environmental sample contained material from six different sites of concentration of adult cattle and/or high traffic areas (e.g. areas surrounding waterers and feeders, areas surrounding the current mobile milking-unit places). Identification of MAP was achieved using a duplex qPCR (Bactotype MAP PCR Kit®, Qiagen). A herd was considered as MAP infected if the environmental sample was positive in the qPCR. Information about the general characteristics of the herd, management practices, and knowledge about the disease was collected using the risk-assessment questionnaire. The information on risk factors was analyzed using a multivariable logistic regression model. The apparent herd-level prevalence was 4.1% (12/292; 95% CI: 1.8-6.4). Herds with a history of mixed farming of cattle with other ruminants had higher odds of being MAP infected than herds without (OR = 3.9; 95% CI: 1.2-13.2). Our study demonstrates the MAP prevalence in dairy herds from Antioquia, Colombia and the possible relationship between MAP environmental positivity with the history of mixed farming of cattle with other susceptible ruminants.

Anisakidae parasites in frozen fish fillets intended for human consumption / Parásitos anisákidos en filetes de pescado congelado destinados a consumo humano

Introduction: Anisakiasis is a human parasitic disease caused by the consumption of raw fish or shellfish containing larvae of the Anisakidae family. It is currently considered an emerging disease of public health interest. Objective: To identify the presence of larvae of the Anisakidae family in samples of frozen raw fish fillets intended for human consumption in markets in Medellín and its metropolitan area in Antioquia, Colombia. Materials and methods: A cross-sectional study was carried out, in which larvae of the Anisakidae family were detected and identified in frozen raw fish fillets from three representative markets in Medellín and its metropolitan area. A total of 384 ready for consumption fillets were analyzed (197 sawfish, 137 salmon, 37 tuna, and 13 hake), using the pressing and ultraviolet light method. Taxonomic keys were used to identify the collected parasites and to establish its genus. Conventional PCR and Sanger sequencing was performed to determine the species. Results: Four larvae were found in 4 of the 384 (1.04%) fillets (CI95% 1.04 ± 1.01%). The species of fish in which the larvae were found was sawfish (Scomberomorus spp.) and the genus and species of the larvae was established as Anisakis pegreffii. Conclusions: According to the study, the presence of Anisakis parasites in frozen raw fish fillets in the influence area is evident.

Introducción. La anisakiasis es una infección producida por parásitos de la familia Anisakidae, transmitida a los humanos por el consumo de pescado o mariscos crudos. En la actualidad, se considera una enfermedad emergente de interés en salud pública. Objetivo. Identificar la presencia de larvas de la familia Anisakidae en muestras de filetes de pescado crudo congelado destinados a consumo humano, en mercados de Medellín y su área metropolitana en Antioquia (Colombia). Materiales y métodos. Se realizó un estudio transversal, en el cual se buscó la presencia de larvas de la familia Anisakidae en filetes de pescado crudo congelado de tres mercados representativos de Medellín y su área metropolitana. Se analizaron 384 filetes listos para el consumo (197 sierras, 137 salmones, 37 atunes y 13 merluzas). Cada filete fue analizado mediante el método de prensado y luz ultravioleta. Los parásitos recolectados se identificaron a partir de claves taxonómicas para establecer el género, así como PCR convencional y posterior secuenciación Sanger, para determinar la especie. Resultados. Se encontraron 4 larvas en 4 de los 384 filetes (1,04 %) (IC95% 1,04 ± 1,01 %). Las larvas encontradas fueron identificadas como Anisakis pegreffi y el tipo de pescado en el cual se encontraron fue la sierra (Scomberomorus spp.) Conclusiones. De acuerdo con el estudio realizado, se evidencia la presencia de parásitos anisákidos en filetes de pescado crudo congelado en el área de influencia.

Characterization of decontamination procedure of beef carcasses at slaughterhouses in the province of Antioquia, Colombia / Caracterización del procedimiento de lavado descontaminación de canales bovinas en las plantas de beneficio animal del departamento de Antioquia, Colombia

Background: Beef slaughterhouses must use a carcass decontamination procedure to control pathogens and thus prevent foodborne diseases transmitted by meat. Objectives: This study aimed to characterize beef carcass decontamination procedures at slaughterhouses located in the province of Antioquia (Colombia). All the slaughterhouses were in service, registered, and approved by Invima (Instituto Nacional de Vigilancia de Medicamentos y Alimentos in Spanish) at the time of the study. Methods: This descriptive study collected information from 23 beef slaughterhouses between July 2019 and April 2021 through documentary reviews and visits to slaughterhouses, using forms and questionnaires. Results: The study allowed the characterization of the procedures used to decontaminate beef carcasses, showing that the chemical disinfection of the carcasses is used to control microorganisms in at least 73.9% of the slaughterhouses analyzed. According to secondary sources, it was found that most of the slaughterhouses are small (slaughter volume <50,000 heads per year); 10 of them use citric acid, lactic acid, peracetic acid, and a mixture of organic acids in concentrations between 900 and 1,200 ppm, 1.5 and 1.7%, 180 and 190 ppm, and 900 and 1,200 ppm, respectively, as carcass disinfectants and according to the technical data sheet of the product. During the visits and through the application of the questionnaire, it was found that the 12 slaughterhouses had implemented chemical disinfection which is not scientifically based, using manual devices as an intervention method to control pathogenic microorganisms. It was found that the type of company, slaughter volume, and the lack of financial resources are the determining factors in the selection of decontamination procedures. The validation of the beef carcass decontamination procedures in the different slaughterhouses in the study was demonstrated. Conclusions:Although it was established that at least one decontamination procedure, such as chemical disinfection, is used in the slaughterhouses of study, this option is not supported by scientific or technical evidence. The findings support the need for improvements in the slaughterhouses of the province of Antioquia, including the improvement of surveillance programs to reduce pathogens in the meat chain effectively.

Antecedentes: Las plantas de beneficio animal deben utilizar un procedimiento de descontaminación de canales para el control de patógenos y con ello, prevenir la aparición de enfermedades transmitidas por el consumo de carne. Objetivos: El objetivo de este estudio fue caracterizar el procedimiento de descontaminación de canales bovinas en las plantas de beneficio animal del Departamento de Antioquia, Colombia, que se encontraban en servicio, inscritas y autorizadas por el Invima al momento del estudio. Métodos: Este estudio descriptivo recolectó información de 23 plantas de beneficio animal de la especie bovina, a partir de revisiones documentales y visitas a las plantas, usando formatos y cuestionarios entre julio de 2019 y abril de 2021. Resultados: El estudio permitió caracterizar los procedimientos y técnicas de descontaminación de canales bovinas, revelando que en al menos el 73,9% de las plantas de beneficio estudiadas se realiza la desinfección química de las canales para el control de microorganismos. A partir de fuentes secundarias, se encontró que la mayoría de las plantas de beneficio animal en el Departamento de Antioquia son muy pequeñas, 10 de ellas utilizan productos de desinfección de canales, tales como el ácido cítrico, ácido láctico, ácido peracético y mezcla de ácidos orgánicos en concentraciones entre 900 y 1200 ppm, 1,5 y 1,7%, 180 y 190 ppm y 900 y 1200 ppm, respectivamente; y estos son utilizados de acuerdo con las recomendaciones de la respectiva ficha técnica del producto. Por otro lado, durante la visita a las plantas de beneficio y mediante la aplicación del cuestionario, se constató que las 12 plantas visitadas han implementado la desinfección química como método de intervención para el control de microorganismos patógenos, realizando su aplicación mediante dispositivos manuales, no obstante, estas prácticas no están fundamentadas científicamente. Por otro lado, se estableció que aparentemente el tipo de empresa, volumen de sacrificio y falta de recursos financieros son los factores que determinan la elección del procedimiento de descontaminación de canales. De igual manera, se evidenció la necesidad de realizar estudios para validar la efectividad del procedimiento de descontaminación en las diferentes plantas de beneficio. Conclusiones: Aunque se estableció que en las plantas de beneficio animal visitadas se implementa al menos una técnica de intervención como la desinfección química, esta elección no tiene un sustento con base a fundamentos científicos y técnicos. Estos hallazgos respaldan la necesidad de mejoras en las plantas de beneficio animal del Departamento, incluyendo mejoras al programa de vigilancia de la reducción efectiva de patógenos en la cadena cárnica.

Detección de anticuerpos anti-Mycobacterium avium subsp. paratuberculosis en bovinos de hatos lecheros de trópico bajo en los departamentos de Sucre y Córdoba, Colombia / Detection of anti-Mycobacterium aviumsubsp. paratuberculosis antibodies in cows of low-tropic dairy herds in the Provinces of Sucre and Córdoba, Colombia

RESUMEN Objetivo. Reportar la frecuencia de la seropositividad a Mycobacterium avium subsp. paratuberculosis (MAP) y explorar los factores asociados al resultado positivo, tanto a nivel de hato como de animal. Materiales y métodos. Se llevó a cabo un estudio transversal en 204 vacas lecheras resultado del cruce de diferentes razas, en cinco hatos de zonas de trópico bajo ubicadas en tres municipios de los departamentos de Sucre y Córdoba (Colombia) en 2018. Los animales fueron seleccionados aleatoriamente y se colectaron muestras de sangre de cada uno. Se utilizó un kit comercial de ELISA para analizar los sueros. La información con respecto a los factores relacionados con los hatos y los animales se recopiló mediante cuestionarios. Se calcularon las estadísticas descriptivas para todas las variables (i.e. tamaño del hato, presencia de otros rumiantes en co-pastoreo con el ganado bovino en los últimos 2 años, edad de los animales, paridad y días desde el último parto), y se exploró la asociación entre dichas variables y el resultado a ELISA (p<0.05). Resultados. El 17.2% (35/204; IC 95%: 12.0-22.3%) de las vacas resultaron positivas a MAP mediante la prueba de ELISA, y los cinco hatos tenían animales seropositivos. La variable a nivel de hato presencia de otros rumiantes en co-pastoreo con el ganado bovino en los últimos 2 años y las variables a nivel de animal edad y paridad se encontraron asociados con los resultados positivos de ELISA. Conclusiones. El presente estudio encontró que el 17.2% de las vacas y el 100% de los hatos fueron positivos a MAP mediante la prueba de ELISA. Adicionalmente se identificaron variables asociadas que pueden ser de interés tanto para los productores como para los veterinarios de animales de producción, y orientar su enfoque para el manejo de la enfermedad.

ABSTRACT Objective. To report the frequency of seropositivity against Mycobacterium avium subsp. paratuberculosis (MAP) and to explore the factors associated with the positive outcome, both at herd and animal level. Materials and methods. A cross-sectional study was carried out on 204 dairy cows, result of the crossing of different breeds, from five low-tropic dairy herds, located in three municipalities of the Provinces of Sucre and Córdoba (Colombia) in 2018. The animals were randomly selected, and blood samples were collected from each one. A commercial ELISA kit was used to analyze the sera. Information regarding factors related to herds and animals was collected through questionnaires. Descriptive statistics were calculated for all variables (i.e. herd size, presence of other ruminants co-grazing with cattle in the last 2 years, age of the animals, parity, and days since the last birth), and the association between these variables and the result to ELISA (p<0.05) was explored. Results. The 17.2% (35/204; 95% CI: 12.0-22.3%) of the cows were positive for MAP by the ELISA test, and the five herds had seropositive animals. The herd-level variable presence of other ruminants in co-grazing with cattle in the last 2 years and the animal-level variables age and parity were associated with the positive ELISA results. Conclusions. The present study found that 17.2% of the cows and 100% of the herds were MAP positive by the ELISA test. Additionally, associated variables were identified and may be of interest to both producers and veterinarians of production animals, guiding their approach to disease management.

Tipificación molecular de Mycobacterium avium subsp. paratuberculosis en lecherías de Antioquia, Colombia

RESUMEN Objetivo. Determinar la diversidad molecular de Mycobacterium avium subsp. paratuberculosis (MAP) en muestras ambientales de hatos lecheros colombianos. Materiales y métodos. Las muestras ambientales de 25 hatos lecheros positivos a MAP por IS900-qPCR se cultivaron por duplicado en medio de yema de huevo de Herrold con micobactina J para obtener aislamientos. Las colonias sospechosas fueron confirmadas para MAP por IS900-qPCR. El ADN positivo se subtipó utilizando técnicas de unidades micobacterialess repetitivas intercaladas - número variable de repeticiones en tándem (MIRU-VNTR) y técnicas de repeticiones de multilocus de secuencia corta (MLSSR) para analizar las diferencias genéticas entre los aislamientos. Resultados. El subtipado reveló dos genotipos diferentes por MIRU-VNTR (INMV 2 e INMV 36). La técnica de MLSSR se realizó para aumentar el poder discriminatorio de lo obtenido por MIRU-VNTR, pero no se observaron diferencias entre los aislamientos recuperados. Conclusiones. El presente estudio representa un enfoque importante para el conocimiento del estatus epidemiológico de MAP en la población de estudio.

ABSTRACT Objective. To determine Mycobacterium avium subsp. paratuberculosis (MAP) molecular diversity in environmental samples from Colombian dairy herds. Materials and methods. Environmental samples from 25 IS900-qPCR MAP-positive dairy herds were cultured by duplicate in Herrold's egg yolk medium with mycobactin J to obtain isolates. Suspicious colonies were confirmed by MAP-IS900-qPCR. Positive DNA was sub-typed using mycobacterial interspersed repetitive units-variable number of tandem repeat (MIRU-VNTR) and multilocus short sequence repeats (MLSSR) techniques to analyze the genetic differences between the isolates. Results. Sub-typing revealed two different genotypes by MIRU-VNTR (INMV 2 and INMV 36). MLSSR technique was carried out to increase the discriminatory power from what was obtained by MIRU-VNTR, but no differences were observed among the recovered isolates. Conclusions. The present study represents an important approach to the knowledge on MAP epidemiological status in the study population.

Detección de anticuerpos anti-Mycobacterium avium subsp. paratuberculosis (MAP) en venados silvestres en Colombia / Detection of anti-Mycobacterium aviumsubsp. paratuberculosis antibodies in wild deer in Colombia

RESUMEN Objetivo. Determinar la presencia de anticuerpos anti-Mycobacterium avium subsp. paratuberculosis (MAP) en venados de cola blanca (Odocoileus virginianus) y venados de páramo (Mazama rufína), capturados en las regiones de la Orinoquía y el Caribe en Colombia. Materiales y métodos. Un total de 44 muestras de suero sanguíneo de venados de las especies M. rufína y O. virginianus fueron colectadas en condiciones de campo entre 2014 y 2016. Se utilizó un kit comercial de ELISA para la detección de anticuerpos anti-MAP. Un animal se consideró positivo a ELISA cuando la relación muestra-positivo (S/P%) fue > 0.4, según lo recomendado por el fabricante. Resultados. El 50% (22/44) de los animales muestreados resultaron positivos, lo que corresponde a 10 hembras y 12 machos. Un 81.8% (18/22) y un 77.3% (17/22) de estos animales seropositivos fueron capturados en la región de la Orinoquía y fueron reportados como adultos, respectivamente. Conclusiones. No se sabe cómo o cuándo se introdujo MAP en la población de ciervos colombianos en las regiones de estudio. La hipótesis más plausible para explicar la presencia de anticuerpos anti-MAP en estas poblaciones silvestres es la transmisión por contacto con el ganado bovino infectado, ya que en ambas regiones estas especies comparten pasturas. Este es el primer estudio en explorar la infección por MAP en animales silvestres en Colombia. Estos hallazgos respaldan la necesidad de realizar más estudios utilizando técnicas de diagnóstico directo, y aproximaciones investigativas que permitan la definición de vínculos en la dinámica de la infección entre mamíferos silvestres y domésticos en Colombia.

ABSTRACT Objective. To determine the presence of anti-Mycobacterium avium subsp. paratuberculosis (MAP) antibodies in white-tailed deer (Odocoileus virginianus) and red brocket deer (Mazama rufína), captured in the Orinoquía and Caribbean regions in Colombia. Materials and methods. A total of 44 blood serum samples from deer of species M. rufina and O. virginianus were collected under field conditions between 2014 and 2016. An ELISA commercial kit was used to detect anti-MAP antibodies. An animal was considered ELISA-positive at a sample-to-positive ratio (S/P%) of > 0.4, as recommended by the manufacturer. Results. The 50% (22/44) of the animals were positive, corresponding to 10 females and 12 males. An 81.8% (18/22) and 77.3% (17/22) of these seropositive animals were captured in the Orinoquía region and were reported as adults, respectively. Conclusions. It is not known how or when MAP was introduced in the Colombian deer population in the study regions. The most plausible hypothesis to explain the presence of antibodies against MAP in these wild populations is transmission by contact with infected bovine cattle since, in both regions, these species share pastures. This is the first study to explore MAP infection in wild animals in Colombia. These findings support the need for further studies using different direct diagnostic techniques and research approaches that allow the definition of links in the infection dynamics between wild and domestic mammals in Colombia.

Tasa de incidencia de mastitis clínica y susceptibilidad antibiótica de patógenos productores de mastitis en ganado lechero del norte de Antioquia, Colombia* / Incidence rate of clinical mastitis and antibiotic susceptibility of mastitis-producing pathogens in dairy cattle from Northern Antioquia, Colombia / Taxa de incidência de mastite clínica e susceptibilidade antibiótica de patógenos produtores de mastite em gado leiteiro do norte de Antioquia, Colômbia

Resumen Introducción: la mastitis se define como la inflamación de la glándula mamaria causante de grandes pérdidas económicas en hatos lecheros, lo que amerita su investigación. Objetivo: determinar la tasa de incidencia de mastitis clínica TIMC, la etiología asociada y la sensibilidad antibiótica de patógenos aislados de casos de mastitis en ganado lechero. Materiales y métodos: se siguieron 37 hatos por 12 meses para el diagnóstico de mastitis cílínica (MC). A las muestras de leche de los casos de MC se les realizó cultivo bacteriológico y a estas, así como a los aislamientos de casos de mastitis subclínica de un estudio previo, se les efectuó prueba de susceptibilidad antibiótica. Para el análisis de la información se utilizó estadística descriptiva estándar. Resultados: TIMC fue de 13,8 casos por 100 vacas-año a riesgo. Se aislaron 188 patógenos totales de casos de MC, entre los cuales el Streptococcus agalactiae, el Streptococcus pyogenes y el Corynebacterium spp. fueron los más frecuentes, con 29,8; 11,7 y 5,9 %, respectivamente. Se halló una alta sensibilidad de las bacterias contagiosas para los antibióticos cloxacilina y cefoperazone. Conclusiones: en el presente estudio se encontró una TIMC de 13,8 casos por 100 vacas-año a riesgo. Los patógenos más prevalentes identificados en casos de MC fueron contagiosos. Se encontró una alta sensibilidad de las bacterias contagiosas para la mayoría de los antibióticos β-lactámicos.

Abstract Introduction: Mastitis is defined as the inflammation of the mammary gland causing great economic losses in dairy herds, which merits investigation. Objective: To determine the incidence rate of clinical mastitis IRCM, associated etiology, and antibiotic sensitivity of pathogens isolated from cases of mastitis in dairy cattle. Materials and methods: 37 herds were followed during 12 months for clinical mastitis (CM) diagnosis. Milk samples from CM cases were bacteriologically cultured, which underwent, along with isolates from subclinical mastitis cases from a previous study, antibiotic susceptibility testing. For data analysis, standard descriptive statistics were used. Results: The IRCM was 13.8 cases per 100 cows-year at risk. A total of 188 pathogens of CM cases were isolated, among which Streptococcus agalactiae, Streptococcus pyogenes, and Corynebacterium spp. were the most frequent, with 29.8%, 11.7%, and 5.9%, respectively. A high susceptibility of contagious bacteria was found for cloxacillin and cefoperazone. Conclusions: The present study found an IRCM of 13.8 cases per 100 cows-year at risk. The most prevalent pathogens identified in cases of CM were contagious. A high sensitivity of contagious bacteria was found for most β-lactam antibiotics.

Resumo Introdução: a mastite se define como a inflamação da glândula mamária causante de grandes perdas econômicas em rebanhos de gado leiteiro, o que justifica a sua pesquisa. Objetivo: determinar a taxa de incidências de mastite clínica TIMC, a etiologia associada e a sensibilidade antibiótica de patógenos isolados de casos de mastite em gado leiteiro. Materiais e métodos: seguiram-se 37 rebanhos bovinos durante 12 meses para o diagnóstico de mastitis clínica (MC). Com as amostras de leite dos casos de MC realizou-se cultivo bacteriológico e a estas, assim como aos isolamentos de casos de mastite subclínica de um estudo prévio, efetuou-se prova de susceptibilidade antibiótica. Para a análise da informação se utilizou estatística descritiva padrão. Resultados: a TIMC foi de 13,8 casos por 100 vacas-ano a risco. Foram isolados 188 patógenos totais de casos de MC, entre os quais o Streptococcus agalactiae, o Streptococcus pyogenes e o Corynebacterium spp. foram os mais frequentes, com 29,8; 11,7 e 5,9 %, respectivamente. Constatou-se uma alta sensibilidade das bactérias contagiosas para os antibióticos cloxacilina e cefoperazone. Conclusões: neste estudo evidenciou-se uma TIMC de 13,8 casos por 100 vacas-ano a risco. Os patógenos mais prevalentes identificados em casos de MC foram contagiosos. Encontrou-se uma alta sensibilidade das bactérias contagiosas para a maioria dos antibióticos β-lactâmicos.

Mycobacterium avium subsp. paratuberculosis in Colombia (1924-2016): A review / Mycobacterium avium subsp. paratuberculosis en Colombia (1924-2016): Revisión de literatura / Mycobacterium avium subsp. paratuberculosis na Colômbia (1924-2016): Revisão de literatura

Abstract Mycobacterium avium subsp. paratuberculosis (MAP) is an acid-fast, gram-positive bacillus. MAP is the causal agent of paratuberculosis (PTB) or Johne's disease, an infectious disease affecting domestic ruminants and some wild species. Its importance as a potentially zoonotic agent due to its relation to Crohn's disease (CD) in humans is still under debate and investigation. The aim of the present systematic review is to summarize original studies on MAP carried out in Colombia since 1924, as well as to establish strengths, weaknesses, and future research opportunities in the country with emphasis on diagnosis and epidemiology. The initial search for existing publications reporting original studies on MAP, PTB, and the relationship between MAP and CD was carried out in the available databases and national libraries. After compilation of the available studies (n = 20), the relevant data was extracted (year, province of report, species studied, diagnostic tests used, study design, summary of results, and authors). Recommendations for future research opportunities on MAP in Colombia are made.

Resumen Mycobacterium avium subsp. paratuberculosis (MAP) es un bacilo ácido resistente, gram-positivo. MAP es el agente causal de la paratuberculosis (PTB) o enfermedad de Johne, una enfermedad infecciosa que afecta rumiantes domésticos y algunas especies salvajes. Su importancia como agente zoonótico, debido a su relación con la enfermedad de Crohn (CD) en humanos, está aún en debate y bajo investigación. El objetivo de la presente revisión es exponer los estudios originales sobre MAP llevados a cabo en Colombia desde 1924, así como establecer sus fortalezas, debilidades y oportunidades de investigación futura, con énfasis en los puntos de vista diagnóstico y epidemiológico. La búsqueda inicial de las publicaciones existentes sobre estudios originales realizados acerca de MAP, PTB y la relación MAP y CD fue realizada en las bases de datos disponibles y en bibliotecas nacionales. Luego de la compilación de los estudios disponibles (n = 20), los datos relevantes fueron extraídos (año, provincia de reporte, especie estudiada, prueba diagnóstica usada, diseño del estudio, resumen de resultados y autores). Se hacen recomendaciones para futuras investigaciónes de MAP en Colombia.

Resumo Mycobacterium avium subsp. paratuberculosis (MAP) é um ácido forte, bacilo gram-positivo. O MAP é o agente causador da paratuberculosis (PTB) ou doença de Johne, uma doença infecciosa que afecta os ruminantes domésticos e algumas espécies selvagens. Sua importância como um agente zoonótico por causa de sua relação com a doença de Crohn (CD) em humanos ainda está em discussão e sob investigação. O objetivo desta revisão é apresentar a estudos mapa original realizado na Colômbia desde 1924, e estabelecer pontos fortes, pontos fracos e oportunidades para futuras pesquisas no país com ênfase no diagnóstico e epidemiológico pontos de vista. A busca inicial da literatura sobre estudos originais sobre MAP, PTB e a relação MAP e CD foi feita nas bases de dados disponíveis e bibliotecas nacionais. Após a compilação dos estudos disponíveis (n = 20), os dados relevantes foram extraídos (ano, relatório província, espécies estudadas, teste de diagnóstico utilizado, desenho do estudo, os resultados resumo e autores). Foram feitas recomendações sobre futuras oportunidades de pesquisa em MAP em Colômbia.

Cross-sectional study of 13 Leptospira serovars in cows in a Colombian dairy region / Estudio transversal de 13 serovares de Leptospira en vacas en una región lechera colombiana / Estudo transversal de 13 sorotipos de Leptospira em vacas em uma região leitera colombiana

Abstract Background: Leptospirosis in cattle is recognized as a major cause of reproductive loses due to abortions, early embryonic death, and infertility. The type of Leptospira serovars that are currently circulating in Colombian herds is largely unknown due to the lack of national reports. Objective: The prevalence of antibodies specific to 13 Leptospira spp serovars was investigated in unvaccinated cows from 26 herds in San Pedro de los Milagros (Province of Antioquia) dairy region. Methods: Microscopic agglutination test was used on serum samples, following WHO recommendations. Results: A total of 112/900 (12.4%) of the animals had antibody titers of 1:100 or greater to at least one Leptospira serovar. Of those positive, 34/900 (3,8%) animals seroconverted to two or more serovars. In order of decreasing prevalence, the serovars detected were: hardjo prajitno, pomona, grippotyphosa, tarassovi, copenhageni, canicola, cellodeni and bratislava. There were 28 animals seropositive to serovar hardjo. Conclusion: Our results indicate that leptospirosis should be included as a differential diagnosis for various reproductive problems in this region.

Resumen Antecedentes: La leptospirosis es la mayor causal de pérdidas reproductivas debidas a abortos, muerte embrionaria temprana e infertilidad. Se desconocen los serovares de Leptospira que actualmente se encuentran circulando en el rebaño colombiano debido a la falta de reportes nacionales. Objetivo: Se investigó la prevalencia de anticuerpos específicos para 13 serovares de Leptospira spp en vacas no vacunadas de 26 rebaños en la región lechera de San Pedro de los Milagros (Provincia de Antioquia). Métodos: La prueba de aglutinación microscópica fue empleada en muestras de suero de acuerdo a las recomendaciones de la OMS. Resultados: Un total de 112/900 (12,4%) de los animales presentaron títulos de anticuerpos de 1:100 o superiores para al menos un serovar de Leptospira. Entre los positivos, el 34/900 (3,8%) de los animales presentaron seroconversión para dos o más serovares. En orden decreciente de prevalencia, los serovares detectados fueron: hardjo prajitno, pomona, grippotyphosa, tarassovi, copenhageni, canicola, cellodeni y Bratislava. En total, 28 animales fueron seropositivos al serovar hardjo. Conclusión: La leptospirosis debería ser incluida como diagnóstico diferencial en problemas reproductivos en esta región.

Resumo Antecedentes: Aleptospirose em bovinos é reconhecida como uma das principais causas de perdas reprodutivas devido a abortos, morte embrionária precoce e infertidade. Os sorotipos de Leptospira que estão atualmente em circulação no rebanho colombiano não se conhecem devido à falta de reportes nacionais. Objetivo: Neste estúdio foi investigada a prevalência de anticorpos específicos para 13 sorovares de Leptospira spp em vacas não vacinadas de 26 rebanhos leiteiros da região de San Pedro, Antioquia. Métodos: Foi empregado o teste de aglutinação microscópica (MAT) em amostras de soro de acordo com as recomendações da OMS. Resultados: Um total de 112/900 (12,4%) animais apresentaram títulos de anticorpos de 1:100 ou superiores para pelo menos um sorotipo de Leptospira. Dos animais positivos, 34/900 (3,8%) soroconverteram para dois ou mais sorotipos. A fim de diminuir a prevalência, foram detectados os sorotipos: hardjo, prajitno, pomona, grippotyphosa, tarassovi, copenhageni, canicola, cellodeni e Bratislava. Um total de28 animais foram soropositivos para o sorotipo hardjo. Conclusão: Os resultados indicam que a leptospirose deveria ser incluída nos diagnósticos diferenciais para o diagnóstico diferencial de problemas em doenças reprodutivas nos bovinos de Antioquia.