DETECCIÓN SEROLÓGICA Y MOLECULAR DE LENTIVIRUS DE PEQUEÑOS RUMIANTES QUE CIRCULAN DE FORMA NATURAL EN OVINOS DE DOS ESTADOS DEL ALTIPLANO MEXICANO.

Los lentivirus de pequeños rumiantes (LvPR) presentan una amplia distribución mundial, provocan enfermedades multisistémicas, crónicas degenerativas e incurables en ovinos y caprinos. En 2010, se realiza la primera secuenciación del genoma completo de LvPR en México y se clasificó dentro del subgrupo B1. Para el 2016, las infecciones por lentivirus en los ovinos fueron incluidas en el Diario Oficial de la Federación. Por lo que los reportes sobre la infección natural por LvPR en ovinos son pocos, el objetivo de este trabajo fue detectar serológica y molecularmente la infección por lentivirus en ovinos y caprinos de dos estados de la región del altiplano mexicano y la evaluación de la capacidad antigénica de una proteína recombinante CAp25. Se encontró la presencia de anticuerpos de ovinos y caprinos contra la proteína SU, gp135, y solo de caprinos contra la CA, p25, el análisis de los epitopes antigénicos se encontraron diferencias antigénicas en las regiones inmunodominantes de la proteína CA, p25 en aislamientos ovinos previamente reportados por otros autores. Aunado a esto, en el presente trabajo, se logró determinar la presencia de provirus tanto en ovinos como en caprinos. Con lo que se podemos concluir que la combinación entre métodos moleculares y serológicos, representa una estrategia importante para la identificación de infecciones por LvPR en los ovinos y caprinos.El estudio fue financiado por el proyecto de investigación: “Desarrollo de una prueba de ELISA indirecta basada en la proteína recombinante de la cápside (CAp25) expresada en E. coli, para el diagnóstico en lentivirus en caprinos”. Recursos Fiscales INIFAP No. SIGI. 1115534018

Prolazna apneja u ovaca: alternativna metoda za uzastopno uzimanje uzoraka mokraće.

Urine samples are necessary in order to carry out many veterinary diagnostic and research protocols. In ovine, these samples are usually obtained through catheterization, but a transitory apnoea could be an easy way to obtain urine samples. This is achieved by preventing air from entering through the nostrils and mouth in sheep, for a short period of time, always less than 1 minute. The results of this transitory apnoea demonstrated that urine characteristics (pH, density and proteinuria) were not different from samples obtained by bladder catheterization. There were no differences in the blood parameters studied (cortisol, glycaemia, total proteins, haematocrit and total and differential leucocyte counts) either, when compared with the control group of catheterized animals, or with the measured values before or after sampling. In conclusion this system could be considered an easy and effective method to obtain urine samples, without causing stress to the sheep and avoiding the alterations to the blood and urinary parameters measured.Prikupljanje uzoraka mokraće nužno je radi provođenja mnogih veterinarskih dijagnostičkih postupaka i znanstvenih istraživanja. Ti se uzorci u ovaca obično uzimaju kateterizacijom. No, prolazna apneja može biti jednostavan način uzimanja uzoraka mokraće u ovaca. To se može postići zatvaranjem nosnica i gubice u kratkom vremenu, uvijek manje od jedne minute. Rezultati pretrage mokraće prikupljene nakon prolazne apneje pokazali su da značajke mokraće (pH, gustoća i proteinurija) nisu bile različite od onih dobivenih pretragom uzoraka uzetih kateterizacijom mokraćnog mjehura. Nije bilo razlike u pretraženim krvnim pokazateljima (kortizolu, glikemiji, ukupnim proteinima, hematokritu, ukupnim leukocitima i diferencijalnoj krvnoj slici) u usporedbi s kontrolnom skupinom kateteriziranih životinja, a ni u usporedbi s izmjerenim vrijednostima prije i nakon uzimanja uzoraka. Može se zaključiti da je opisani način uzimanja uzoraka jednostavan i učinkovit bez izazivanja stresa u ovaca i ne dovodi do promjena u pokazateljima krvi i mokraće

Inducción experimental de epididimitis en ovinos por inoculación intrauretral con Actinobacillus seminis: estudio bacteriológico, serológico e histopatológico

Actinobacillus seminis infection was experimentally induced in 18 6-month-old lambs. Four animals were kept as negative controls, three animals were inoculated with A. seminis by the intra-epididymal (IE) route, and 11 lambs were inoculated by the intra-urethral (IU) route, as follows: Five days prior to challenge, 4 lambs were given gonadotropin-releasing factor (GnRH), 4 lambs received ethylene glycol IE 24 h prior to challenge, and 3 lambs received no previous treatment. Two of the 4 negative controls received GnRH, and 2 received ethylene glycol. Serum samples were collected weekly for immunodiffusion. Similarly, semen samples were collected for bacteriological analysis. Animals were slaughtered 35 d postinoculation. Testis, epididymis, and accessory sex gland samples were collected for bacteriological and histopathological analyses. A. seminis was isolated from the semen of the GnRH-treated group, but not from the reproductive tract. Lesions had been previously observed in 3 of these 4 animals. A. seminis was isolated from both the ampulla ductus deferens and from the seminal vesicle of one ethylene glycol-treated animal. A. seminis was isolated from the semen of 2 of the 3 lambs that were challenged with no previous treatment. A. seminis was isolated from the inoculation site of one lamb and from the accessory sex glands of 2 of the 3 IE-inoculated lambs. Two of the 11 IU-inoculated lambs showed positive serological results. Two of the 3 IE-inoculated lambs had positive serological results. Histopathological lesions were consistent with epididymitis, including inflammatory infiltrates in the accessory glands. Results suggest that the accessory sex glands can be an A. seminis reservoir.Con el objetivo de inducir la infección experimental de A. seminis, se utilizaron 18 corderos de seis meses de edad: 4 testigos negativos, otros 3 se inocularon con A. seminis vía intraepididimal (IE) y 11 fueron inoculados por vía intrauretral (IU) como sigue: cuatro recibieron factor liberador de gonadotropinas (GnRH) cinco días previos al desafío, cuatro recibieron etilenglicol por vía IE 24 h previas al desafío y tres no recibieron tratamiento previo. De los cuatro testigos negativos, dos recibieron GnRH y dos etilenglicol. Se tomaron muestras semanales de suero para inmunodifusión, y de semen para bacteriología. Se sacrificaron a los 35 días posinoculación, obteniéndose testículos, epidídimos y glándulas anexas, para estudios bacteriológicos e histopatológicos. Del grupo tratado con GnRH, se recuperó la bacteria del semen pero no del aparato reproductor, aunque se observaron lesiones en 3 de 4 animales. Del grupo estimulado con etilenglicol, en uno se aisló A. seminis de ámpulas del deferente y de vesícula seminal. De los desafiados sin tratamiento previo, en 2 de 3 se aisló la bacteria del semen. De los tres inoculados IE, en uno se aisló del lugar de inoculación y en dos se aisló de una de las glándulas anexas. En la serología de los desafiados por vía IU, 2 de 11 resultaron positivos, de los 3 inoculados IE, 2 resultaron positivos. Las observaciones histopatológicas correspondieron a lesiones típicas de epididimitis con infiltrados inflamatorios en glándulas anexas. Los resultados sugieren que las glándulas anexas pueden ser un reservorio de A.seminis

First identification and characterization of Streptococcus iniae obtained from tilapia (Oreochromis aureus) farmed in Mexico

Estudio de cuadros septicémicos por Streptococcus iniae en tilapias de cultivo de México.This is the first study to isolate, identify and characterize Streptococcus iniae as the causative disease agent in two tilapia (Oreochromis aureus) populations. The populations were geographically isolated, of distinct origins, and did not share water sources. Affected fish showed various external (e.g., exophthalmia and cachexia, among others) and internal (e.g., granulomatous septicaemia and interstitial nephritis, among others) signs. All internal organ samples produced pure cultures, two of which (one from each farm, termed S-1 and S-2) were subjected to biochemical, PCR and 16S rRNA sequencing (99.5% similarity) analyses, confirming S. iniae identification. The two isolates presented genetic homogeneity regardless of technique (i.e., RAPD, REP-PCR and ERIC-PCR analyses). Pathogenic potentials were assessed through intraperitoneal injection challenges in rainbow trout (Oncorhynchus mykiss) and zebrafish (Danio rerio). Rainbow trout mortalities were respectively 40% and 70% at 104 and 106 CFU per fish with the S-1 isolate, while 100% mortality rates were recorded in zebrafish at 102 and 104 CFU per fish with the S-2 isolate. The obtained data clearly indicate a relationship between intensified aquaculture activities in Mexico and new disease appearances. Future studies should establish clinical significances for the tilapia industry.Universidad Autónoma del Estado de México, Grant/Award Number: 3675/2014/CID and 4489/2018/CI. Comisión Nacional de Investigación Científica y Tecnológica, Grant/Award Number: FONDAP 15110027, FONDECYT 115069

CARACTERIZACIÓN DE Escherichia coli AISLADOS DE HISOPADOS RECTALES DE CORDEROS DEL ESTADO DE MÉXICO.

E. coli es uno de los microorganismos más difundidos en la naturaleza y además el integrante principal de la flora intestinal normal, pero consistentemente la bacteria es uno de los agentes etiológicos más importantes de las diarreas neonatales de los mamíferos, concretamente cuando intervienen determinadas cepas de E. coli que poseen una peculiar capacidad de adhesión selectiva de las células epiteliales del intestino (Gallego y Pérez, 1993). Las diferentes investigaciones realizadas señalan que E. coli está presente en el 30% al 40% de los focos de diarrea neonatal de los corderos, pero en concordancia con el principio etiológico del síndrome diarreico neonatal, la presentación de la diarrea colibacilar, al igual que la producida por otros microorganismos, no va a depender en exclusiva de las complejas características antigénicas, serológicas y de los factores de virulencia de las cepas de E. coli. La edad del cordero, con una mayor susceptibilidad cuanto más temprana, y el nivel de inmunidad pasiva aportado por el calostro, son igualmente determinantes, asimismo, varios de los elementos del factor entorno van a condicionar la presentación de la enfermedad; en concreto los aspectos que se engloban en lo que habitualmente se denomina manejo, como deficiencias en higiene, humedad, temperatura ambiente, hacinamiento y coexistencia de edades muy diferentes (Gallego y Pérez 1993). Las infecciones sistémicas causadas por E. coli son bastantes frecuentes en terneros, corderos y pollos. Las cepas septicémicas de E. coli presentan mecanismos especiales para superar las defensas del hospedador. Son capaces de alcanzar la corriente sanguínea tras penetrar por el intestino, los pulmones o el tejido umbilical (Quinn et al., 2005).E. coli es uno de los microorganismos más difundidos en la naturaleza y uno de los integrantes principales de la flora intestinal normal, pero algunos serotipos son agentes causales de diarreas neonatales en los mamíferos. Las diferentes investigaciones realizadas señalan que E. coli está presente en el 30% al 40% de los focos de diarrea neonatal de los corderos. La resistencia a los antimicrobianos es un problema importante que va en aumento en humanos como en animales. El amplio uso y a veces indiscriminado de estos compuestos da lugar a una selección de bacterias que son hereditariamente resistentes. El objetivo de este trabajo es evaluar la resistencia a los antibióticos β-lactámicos, quinolonas, tetraciclinas y aminoglucósidos en cepas de Escherichia coli aisladas de corderos con y sin diarrea y madres. Se tomaron 182 muestras de hisopados rectales en ocho unidades de producción del estado de México, se realizó la siembra en agar EMB para el aislamiento de E. coli, a las colonias sospechosas se les realizaron pruebas bioquímicas para su identificació

Phenotypic and genotypic profile of clinical and animal multidrug-resistant Salmonella enterica isolates from Mexico

Descripción de Salmonella enterica multirresistenteAims: The objective of this study was to obtain a phenotypic and genotypic profile of Salmonella enterica including multidrug-resistant (MDR) isolates from food-producing animals and clinical isolates, as well as their genetic relatedness in two different States of Mexico (Jalisco and State of Mexico). Methods and Results: A total of 243 isolates were evaluated in terms of antimicrobial resistance (AMR) and related genes through a disk diffusion method and PCR respectively; we found 16 MDR isolates, all of them harbouring the blaCMY gene but not qnr genes, these isolates represent less than 10% of the collection. The pulsed-field gel electrophoresis revealed a higher genotypic similitude within isolates of State of Mexico than Jalisco. Conclusions: A low percentage of Salmonella isolates were resistant to relevant antibiotics in human health, nevertheless, the AMR and involved genes were similar despite the different serovars and origin of the isolates. Significance and Impact of the Study: This investigation rovided an insight of the current status of AMR of Salmonella isolates in two States of Mexico and pinpoint the genes involved in AMR and their epidemiological relationship, the information could help to determine an adequate therapy in human and eterinary medicine

Serotypes, virulence genes profiles and antimicrobial resistance patterns of Escherichia coli recovered from feces of healthy lambs in Mexico

Articulo que habla de la resistencia a los antibióticos en corderosHealthy lambs are one of the major reservoirs of Shiga toxin-producing Escherichia coli (STEC) and it is known as the cause of foodborne diseases (FBD). The work objective is to characterize (STEC) isolates obtained from rectal swabs of healthy lambs herds, a total of 183 samples were obtained from sheep production units of the State of Mexico. E. coli isolates were confirmed through the amplification of the uid A gene. antimicrobial sensitivity pattern was determined through Kirby-Bauer (CLSI, 2012) test and the presence stx1, stx2 and eae genes from isolates by multiplex PCR. Serotyping was performed using specific anti-O and anti-H sera (SERUNAM, Mexico) for 185 Somatic and 56 flagellar antigens. 126 isolates biochemically and molecularly identified as E. coli were obtained, of which 80 did not express any virulence factor and 46 expressed at least some (STEC) virulence factor. The highest percentage of E. coli resistance was for tetracycline 48.7% (39/80), followed by nalidixic acid 13.7% (11/80), gentamicin 6.2% (5/80) and Ciprofloxacin 3.7% (3/80). Resistance to amikacin, cefotaxime and ceftazidime were not detected. A frequency of 46 STEC isolates (36.2%) were obtained, of which 28/46 (22.0%) expressed stx1, stx2 3/46 (6.5%), stx1, stx2 13/46 (10.2%) and eae 2/46 (1.6%). Thirty different serotypes were obtained. The three serotypes with the highest number of isolates (four each) were: O76:H19, O118:H27 and O146:H21 which have been identified as a cause of diarrhea in human population. An isolate of serogroup O104 was obtained, with a significant importance for European public health. In virtue of the discovered serotypes and the virulence factors distribution, we can affirm that the obtained isolates from lambs in the State of Mexico are classifiable as atypical STEC of low virulence

Effects of dietary chromium-yeast level on growth performance, blood metabolites, meat traits and muscle fatty acids profile, and microminerals content in liver and bone of lambs

To assess the effect of dietary supplement levels of chromium-yeast (Cr-yeast) on growth performance, blood glucose and triglycerides, fatty acid (FA) profile in intramuscular fat, carcase and meat traits, iron, copper, chromium and zinc concentrations in liver and bone, 24 Rambouillet male lambs (29.2 ± 0.17 kg body weight) were randomly assigned to four diets with 0, 0.2, 0.4 and 0.6mg Cr/kg DM. The growth performance trial lasted 49 d. Supplemental Cryeast did not affect growth performance and carcase characteristics (p>.05), but reduced (p<.05) perirenal and intramuscular fat, as well as 3 h post-feeding blood glucose and triglycerides concentration. In liver, Fe and Cu concentration decreased (p<.05), while Cr concentrations in liver increased with increasing Cr-yeast dietary levels. In bone, Fe decreased (p<.05) as Cryeast dietary levels increasing, and Cr-yeast supplementation increased Cr concentrations (p<.05). As Cr-yeast dietary level increased, palmitic (C16:0) and stearic (C18:0) SFA decreased linearly (p<.05), while palmitoleic (C16:1n-7), vaccenic (C18:1n-7), linoleic (C18:2n-6) and arachidic (C20:4) unsaturated fatty acids (UFA) increased linearly (p<.01). In conclusion, Cr-yeast did not affect growth performance and carcase quality, but decreased the perirenal and intramuscular fat, blood glucose and triglyceride content, and Fe and Cu concentrations in liver as increased Cr-yeast levels in the diet. Because supplemental Cr-yeast improved index of atherogenicity and unsaturated to saturated FA ratio in muscle of lambs, it could be of human nutritional interest

Effects of Ewe’s Diet Supplementation with Polyunsaturated Fatty Acids on Meat Lipid Profile of Suckling Lambs

[EN] Polyunsaturated fatty acid (PUFA) deposition in lambs’ muscles could be influenced by their mothers’ diet. The aim was to study the profile of fatty acids in the muscle of lambs from ewes supplemented with different sources of PUFA to achieve a healthier meat for the consumer. On day 100 of gestation, pregnant ewes grazed on natural grass were divided into three groups (n = 20) and supplemented with PUFA: Group A: 700 g of a ration rich in PUFA, Group B: 700 g of a ration for sheep + 20 mL of fish oil and Group C: 700 g of the same ration. After parturition, each group was subdivided: ten ewes continued with the same diet until the end of lactation; the other ten were fed only natural grass. The values of n-3 and n-6 PUFA and alpha-linolenic acid (ALA), eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), linoleic acid (LA) and arachidonic acid (AA) were analyzed in Longissimus lumborum muscles of lambs at 90 and 120 days of life. The feeding of ewes during lactation favorably influenced the lipid profile of the lamb muscle, increasing the concentration of n-3 and n-6 PUFA, ALA, LA, AA, DHA and EPA. The supplementation of ewes with fish oil and/or a ration rich in PUFA improved the LA/ALA ratio in lambs’ meatSIThis research was funded by the CSIC-UdelaR (Comisión Sectorial de Investigación Científica, Universidad de la República), project “Metabolic, behavioral parameters and concentration of fatty acids in carcass of lambs born to mothers supplemented with long-chain polyunsaturated fatty acids”. 201

Aislamiento e identificación fenotípica y genotípica de Moraxella ovis de casos clínicos de queratoconjuntivitis ovina en el Estado de México

La queratoconjuntivitis contagiosa ovina (QCO) es una enfermedad infectocontagiosa que produce ceguera temporal o permanente en ovinos y caprinos, se encuentra asociada a un conjunto de agentes infecciosos como Moraxella ovis, Mycoplasma conjunctivae y Chlamydia psittaci, el diagnóstico se realiza mediante un examen clínico y pruebas de laboratorio. De un total de 861 animales examinados, 209 presentaron algún tipo de lesión ocular resultando en una prevalencia del 24.27% de animales con lesiones compatibles con QCO. De las 209 muestras remitidas al laboratorio se lograron identificar 58 como Moraxella ovis mediante bacteriología y por la amplificación de los genes 16S rRNA y RxtA por PCR. En virtud de los resultados podemos concluir que Moraxella ovis esta involucrada en los casos de QCO en establecimientos productores de ovinos en el Estado de México