ABSTRACT The objective of this study was to determine the impact of metabolic changes occurring in induced subclinical gestation toxaemia in ewes on metabolic parameters and vigour in the first 72 h of their lambs’ life. Fifty-one adult Corriedale ewes of known gestation date and fed on a natural pasture were randomly divided into four groups at day 145 of gestation. Group A (ewes with single pregnancies) and group C (ewes with twin pregnancies) were fed on a natural pasture throughout the trial; group B (ewes with single pregnancies) and group D (ewes with twin pregnancies) were subjected to 75% feed restriction for three days until they reached glycaemia and β-hydroxybutyrate (BHB) values indicative of subclinical gestational toxaemia. The birth-to-first station and birth-to-first suckling times of the born lambs were studied. Within the first hour after parturition and at 24, 48, and 72 h, glycaemia and BHB concentrations were determined in the blood of lambs, and body temperature and body weight were assessed at the same time. Relative weight gain at 72 h of age was calculated for all lambs. The BHB concentration determined at all times studied showed no difference among the groups. Subclinical gestation toxaemia induced in ewes subjected to feed restriction causes a decrease in the glycaemia of their lambs in the first hour of life; however, it does not cause an increase in ketone bodies. The metabolic changes induced in the mothers do not have negative effects on the vigour of the lambs at birth, as the weight, temperature, and the time it took for lambs to stand and suckle are not affected. However, they have a negative effect on relative weight gain in the critical period of the first 72 h of life. 7 life Fiftyone Fifty one 14 pregnancies trial 75 βhydroxybutyrate β hydroxybutyrate (BHB birthtofirst birth 24 48 however bodies affected However 1 2 4

Abstract Antibiotic resistance genes (ARG) have been described mainly in bacteria, but are known to occur in temperate phages. Prevalence of the qnrB, qnrA and bla TEM genes was identified in Escherichia coli strains and temperate phages by lytic cycle induction. From a total of 48 samples collected from drinking water, wastewater and sewer water in slaughterhouses in the State of Mexico, Mexico, 37 contained E. coli isolates. Resistance was highest to tetracycline (32/37; 86.4 %), followed by trimethoprim-sulfamethoxazole (19/37; 51.3 %) and ampicillin and nalidixic acid (18/37; 48.6 %). Prevalence of the bla TEM gene was 37.8 % in the bacterial isolates and 3.5 % in the phage isolates. The bacterial isolates contained 8.1 % qnrA and 29.7 % qnrB genes, while the phage isolates contained 2.7 and 24.3 %, respectively. Presence of ARG in the bacterial isolates was linked to phage DNA, highlighting the significant role it plays in the spread of ARG in the studied slaughterhouses. Understanding the mechanisms of antimicrobial resistance will contribute to developing effective control measures.

Resumen Los genes de resistencia a los antibióticos (ARG) han sido descritos principalmente en bacterias; sin embargo, en fagos atemperados los estudios han sido escasos. En este estudio se determinó la prevalencia de los genes qnrB, qnrA y bla TEM en cepas de Escherichia coli y en fagos atemperados obtenidos por inducción del ciclo lítico en dichos aislamientos. Se recolectaron 48 muestras de agua potable, agua residual y alcantarillado en rastros del Estado de México obteniendo 37 aislamientos de E. coli. La mayor resistencia fue para tetraciclina 32/37 (86.4 %), seguido de trimetoprim-sulfametoxasol 19/37 (51.3 %) y por último ampicilina y ácido nalidíxico con el mismo número de aislamientos 18/37 (48.6 %). La prevalencia del gen bla TEM en aislamientos bacterianos fue 37.8 %, mientras que en los aislamientos fágicos fue 3.5 %. Los genes qnrA y qnrB fueron encontrado s en 8.1 % y 29.7 % respectivamente en aislamientos bacterianos, mientras que en los aislamientos fágicos fueron obtenidos 2.7 y 24.3 % respectivamente. Los resultados muestran que los ARG presentes en aislamientos bacterianos se relacionan en el ADN fágico, lo que indica el papel significativo en la propagación de ARG en los rastros estudiados. Comprender los mecanismos de resistencia a antimicrobianos, permitirá establecer medidas efectivas para disminuir este fenómeno.

ABSTRACT The objective of this work was to determine the repercussion of the metabolic changes that occur in induced subclinical pregnancy toxemia in single- or twin-bearing ewes on the duration of gestation, type and length of lambing, and placental expulsion time. Fifty-one adult Corriedale ewes with a known gestation date and fed in the wild were randomly divided at day 145 of gestation into four groups. Group A included single-bearing ewes, and C twin-bearing ewes, all of which were fed ad libitum . Group B was composed of single-bearing ewes, and D of twin-bearing ewes, all subjected to a 75% feed restriction until glycemic values were compatible with subclinical pregnancy toxemia. Metabolic parameters were determined in all ewes from day 145 of gestation to 72 h postpartum. Likewise, the duration of gestation, type of lambing (eutocic/dystocic), labor length, and placental expulsion time were recorded. Glycemia and β-hydroxybutyrate values compatible with subclinical pregnancy toxemia were achieved after 48 h of feed restriction, being 30.67±2.37 mg/dL and 1.87±0.12 mmol/L in single-bearing ewes and 28.40±3.39 mg/dL and 2.21±0.42 mmol/L in twin-bearing ewes, respectively. It is concluded that subclinical pregnancy toxemia induced by feed restriction at the end of gestation, frequent situation that occurs in flock management, produces mild metabolic changes, which return to normal values after the delivery. These metabolic changes registered before lambing do not modify the gestation length, do not increase the percentage of dystocical deliveries, nor influence labor length or placental expulsion time.

The objective of this study was to determine the blood distribution of CD4, CD8, and T γδ lymphocytes (WC1) in rams experimentally infected with Brucella ovis. Eighteen rams from 1 to 4 years old and free from B. ovis were distributed in three groups: Control (n=6); Inoculated in the ocular and preputial mucosa (n=6); Inoculated intravenously (n=6). Serological follow-up was carried out from the day of inoculation until 189 dPI (post-inoculation day). CD4 and CD8 lymphocyte populations were immunotyped at 120, 150 and 189 and WC1 (Tγδ lymphocytes) at 120 dPI by flow cytometry. The rams, from the third dIP began to seroconvert; at 21 and 28 dPI all animals of the inoculated intravenously and in the mucosa groups, respectively, were positive; likewise, 50% of the animals of the challenged groups were presented as positive or suspect to the ELISA test at 189 dPI. Differences were found in the Mean Fluorescence Intensity (MFI) of the CD8 lymphocytes between the control group (1027.4) and the group inoculated intravenously (499.6) at 120 dPI (p<0.05) and in the MFI of the lymphocytes CD4 between groups at 189 dPI (p<0.05). The T γδ lymphocyte populations (WC1) presented differences in MFI between the control group and the groups inoculated at 120 dPI (p<0.05). The results indicated that B. ovis can modulate host immune response, that CD4 and CD8 lymphocytes are important for host defence against this infection, and that populations of CD4 and CD8 can fluctuate during the period of B. ovis infection. Besides, the participation of T γδ lymphocytes could be an important factor in the control of the infection caused by B. ovis

El objetivo de este estudio fue determinar la distribución sanguínea de linfocitos T CD4, CD8, LT γδ (WC1) en carneros infectados experimentalmente con Brucella ovis. Se utilizaron 18 carneros de 1 a 4 años y libres de B. ovis distribuidos en tres grupos: Control (n=6); Inoculado en las mucosas ocular y prepucial (n=6); Inoculados vía endovenosa (n=6). Se realizó el seguimiento serológico desde el día de la inoculación hasta el 189 dPI (día pos-inoculación). Se inmunotipificaron las poblaciones de linfocitos CD4 y CD8 a los 120, 150 y 189 y de WC1 (linfocitos Tγδ) a los 120 dPI por citometría de flujo. Los carneros, a partir del tercer dPI comenzaron a seroconvertir; a los 21 y 28 dPI todos los animales de los grupos inoculados por vía endovenosa y mucosa, respectivamente, resultaron positivos; asimismo, el 50% de los animales de los grupos desafiados se presentaron como positivos o sospechosos a la prueba de ELISA a los 189 dPI. Se encontraron diferencias en las Medias de Intensidad de Fluorescencia (MIF) de los linfocitos CD8 entre el grupo control (1027.4) y el grupo inoculado por vía endovenosa (499.6) a los 120 dPI (p<0.05) y en las MIF de los linfocitos CD4 entre grupos a los 189 dPI (p<0.05). Las poblaciones linfocitarias T γδ (WC1) presentaron diferencias en MIF entre el grupo control y los grupos inoculados a los 120 dPI (p<0.05). Los resultados indican que B. ovis puede modular la respuesta inmune del hospedero, que los linfocitos CD4 y CD8 son importantes para la defensa del hospedero ante esta infección, y que las poblaciones de CD4 como de CD8 pueden fluctuar durante el periodo de infección por B. ovis. Así mismo, la participación de los linfocitos T γδ podría ser un factor importante en el control de la infección causada por B. ovis

Resumo: A distribuição das células envolvidas na resposta imune em glândulas sexuais acessórias de carneiros experimentalmente infectados com Actinobacillus seminis foi estudada. Doze carneiros de um ano de idade foram experimentalmente infectados via intrauretral (IU) (n=4) e via intraepididimal (IE) (n=4) e quatro animais controles (CON) foram utilizados. Os animais foram abatidos 35 dias após a inoculação, amostras foram retiradas das glândulas sexuais acessórias e testes bacteriológicos e histopatológicos foram realizados. A presença de linfócitos CD4, CD8 e TCRγδ (WC1), células CD45RO, macrófagos (CD14), células dendríticas (CD1b) e células contendo IgA, IgG and IgM (IgCC) foi determinada. Os animais do grupo IE desenvolveram epididimite clínica. Não foram visualizadas lesões nos carneiros do grupo IU, dois dos CON inoculados intraepididimalmente desenvolveram pequenas lesões no epidídimo. Isolados de A. seminis foram obtidos de 6:16 (37,5%) nas glândulas sexuais acessórias no grupo IE mas não nos grupos IU e CON. No grupo CON células contendo IgA and IgM predominaram nas glândulas bulbouretrais e na próstata e foram escassas ou ausentes nas vesículas e na ampola. Um incremento significativo de células contendo IgA, IgG and IgM foi confirmado nas vesículas seminais, na ampola e nas glândulas bulbouretrais no grupo IE. Nos grupos IE e IU foi evidenciado um aumento em CD4, CD8, WC1, CD45RO e CD14 nas vesículas e ampola. As células dendríticas CD1b estavam presentes na ampola e nas vesículas com processo inflamatório. A. seminis induziu uma resposta imune local nos grupos IE e IU. Estes resultados indicam um padrão diferente de células imunes infiltrantes nas glândulas sexuais acessórias de carneiros infectados por A. seminis.

Abstract: The distribution of cells involved in the immune response in accessory sex glands of rams experimentally infected with Actinobacillus seminis was studied. Twelve one-year old rams were experimentally infected by intraurethral (IU) (n=4) and intraepididymal (IE) (n=4) route, and four control (CON) animals were used. The animals were slaughtered 35 days post-inoculation, samples were taken from accessory sex glands, and bacteriology and histopathology tests were performed. The presence of CD4, CD8 and TCRγδ (WC1) lymphocytes, CD45RO cells, macrophages (CD14), dendritic cells (CD1b), IgA-, IgG- and IgM-containing cells (IgCC) was determined. Animals of the IE group developed clinical epididymitis. No lesions were seen in rams of the IU group; two of the intraepididymal inoculated CON developed small lesions in the epididymis. A. seminis isolates were achieved from 6:16 (37.5%) accessory sex glands in the IE group, but not in the IU and CON groups. In the CON group, IgA- and IgM- containing cells predominated in the bulbourethral glands and the disseminated prostate, and they were scarce or null in the vesicles and ampullae. A significant increase of IgA-, IgG- and IgM- containing cells was confirmed in the seminal vesicles, the ampullae and the bulbourethral glands in the IE group. In the IE and IU groups, an increase in CD4, CD8, WC1, CD45RO and CD14 was evidenced in the vesicles and ampullae. CD1b dendritic cells were present in the ampullae and vesicles with inflammatory processes. A. seminis triggered a local immune response in the IE and IU groups. These results indicate a different pattern of infiltrating immune cells in the accessory sex glands of infected A. seminis rams.

In vitro phagocytosis and apoptosis activity was evaluated in polymorphonuclear leukocytes (PMN) dairy cattle in the presence of Staphylococcus aureus capsular types 5 and 8. PMN were obtained from clinically healthy cows. The phagocytic activity of neutrophils was assayed through the phagocytic capacity and phagocytic index. Apoptosis was determined by light microscopy (LM) and epifluorescence (FM). Differences were observed between treatments in LM regarding the capacity of phagocytosis; compact type (CC) were 86.50±2.93, capsular serotype 5 (CP5) 74.98±2.44 and capsular serotype 8 (CP8) 82.22±1.50 (P<0.05). The values for the phagocytic index were: CC 6.13±0.31, CP5 4.88±0.13 and CP8 5.22±0.10 (P<0.05). The phagocytic capacity in the FM was: CC 86.77±2.6, CP5 62.52±3.26 and CP8 78.60±4.07. The phagocytic index for CC was 7.62±0.55, CP5 4.67±0.29 and CP8 5.53±0.40 (P<0.05). The percentages of apoptosis evaluated by LM were: positive control (C+) 97.7±1.28, negative control (C-) 27.13±1.85, CC 35.44±2.56 CP5 60.07±4.21, CP8 45.57±4.34 and CP5 52.78±3.29. In the FM technique were: C(+) 95.61±2.66, C(-) 20.2±1.77, CC 26.5±1.65, CP8 41.33±2.32 respectively (P<0.05). Differences in the in vitro phagocytosis capacity, phagocytic index and apoptosis of PMN were found between the S. aureus serotypes tested. Type CP5 decreased phagocytic activity and significantly increased the apoptosis compared with type CP8 in PMN.

Se evaluó la actividad de fagocitosis y apoptosis in vitro en leucocitos polimorfonucleares (PMN) de bovinos lecheros frente a Staphylococcus aureus tipos capsulares 5 y 8. Los neutrófilos se obtuvieron de vacas clínicamente sanas. La actividad de fagocitosis de neutrófilos se determinó a partir de la capacidad e índice de fagocitosis. La evaluación de la apoptosis se realizó por microscopía de campo claro (ML) y de epifluorescencia (MF). Se observaron diferencias entre los tratamientos en ML para la capacidad de fagocitosis, el tipo compacto (CC) fue 86.50±2.93, serotipo capsular 5 (CP5) 74.98±2.44 y serotipo capsular 8 (CP8) 82.22±1.5 (P<0.05). Los valores para el índice de fagocitosis fueron: CC 6.13±0.31, CP5 4.88±0.13 y CP8 5.22±0.10 (P<0.05). La capacidad de fagocitosis observada en la MF fue: CC 86.77±2.6, CP5 62.52±3.26 y CP8 78.60±4.07. El índice de fagocitosis para la CC fue de 7.62±0.55, 4.67±0.29 para CP5 y de 5.53±0.40 para CP8 (P<0.05). Los porcentajes de apoptosis por ML fueron: testigo positivo (C+) 97.7±1.28, testigo negativo (C-) 27.13±1.85, CC 35.44±2.56, CP5 60.07±4.21 y CP8 45.57±4.34 (P<0.05). En la técnica de MF fueron: C(+) 95.61±2.66, C(-) 20.2±1.77, CC 26.5±1.65, CP5 52.78±3.29 y CP8 41.33±2.32 (P<0.05). Se encontraron diferencias en la capacidad e índice de fagocitosis y apoptosis de los PMN entre los tipos capsulares de S. aureus evaluados. El CP5 al reducir la actividad de fagocitosis e incrementar la inducción de la apoptosis en los neutrófilos, puede contribuir a la persistencia de la infección intraglandular mamaria del ganado lechero en la mastitis bovina.

Os patógenos do aparelho reprodutor do macho podem penetrar e se estabelecer por via ascendente, a partir do prepúcio à uretra, glándulas anexas, epidídimo e testículos. Neste trabalho foi quantificada a distribuição de algumas das células envolvidas na resposta imune, em nível de prepúcio e uretra pélvica, em quatro carneiros de um ano de idade, sem lesões aparentes no testículo, no epidídimo e no prepúcio. No momento da eutanásia foram coletadas amostras do prepúcio e da uretra pélvica, as quais foram conservadas em formalina 10% e congeladas. De acordó com as análises de imunohistoquímica foram observadas células CD4, CD8, WC1, CD45RO, CD14 e CD1b; e por imunoflorescência foram observadas células que continham imunoglobulinas (CCI) das clases IgA, IgG e IgM. Com um analisador de imagens foi feita a contagem das células marcadas, presentes na mucosa de ambos os orgãos. Foi realizada a comparação entre o total de células entre ambos os tecidos e de forma diferencial entre os tecidos epitelial e conectivo da mucosa. Foram encontradas diferenças significativas no número total de linfócitos CD4, CD45RO e WC1, no número de macrófagos CD14 e células dendríticas CD1b, com valores de médias que foram maiores em todas as amostras coletadas do prepucio, em relação as amostras coletadas da uretra (p<0.05). Células dendríticas só foram encontradas no prepúcio. Não foram encontradas diferenças significativas no número de linfocitos CD8 entre ambos os orgãos. A relação entre cada tipo celular no tecido conectivo e intra-epitelial e entre os diferentes órgãos, resultou para CD4 10/1 no prepúcio (p<0.05), contra 7/1 na uretra (p<0.05), entretanto os CD8 se distribuíram 1/1 em ambas as mucosas, não sendo significativa as diferenças. Os WC1 foram observados na relação 5/1 em ambas as mucosas (p<0.05). A célula CD45RO, no prepucio, foi observada de 19/1(p<0.05) e na uretra de 1/1, não sendo um resultado significativo. As CC-IgA não mostraram diferença significativa no total de células em ambos os tecidos. Na uretra não foram observadas as CC-IgG, e as CC-IgM foram escassas; em contrapartida, ambos os tipos celulares foram observadas no prepucio, em quantidades menores que na uretra (p<0.05). As CC-IgA, IgG e IgM foram observadas em ambos os tecidos conectivos da mucosa. A presença de células apresentadoras de antígenos, macrófagoss e células dendríticas, assim como de linfócitos CD4, CD8. TCR γδ (WC1), CC-IgA, IgG e IgM e células CD45RO, determinam que ambas as mucosas podem se comportar como locais de indução e promoção da resposta imune das mucosas.

The pathogens of the reproductive system in the male can penetrate and establish by ascending route, from to the prepuce to the urethra, accessory glands, epididymis and testicles. The aim of this paper is determine the distribution and number of cells involved in the immune response in prepuce and pelvic urethra of rams, without apparent clinical alterations in testicle, epididymis and prepuce. The distribution of some of the cells involved in the immune response at the level of the prepuce and the pelvic urethra was quantified in four one-year-old rams seronegative for B. ovis and A. seminis and without apparent lesions in the testicles, the epididymis, and the prepuce. At the moment of slaughter, samples were taken from the preputial fornix and the pelvic urethra and placed in 10% formalin and under freezing conditions. CD4, CD8, WC1, CD45RO, CD14 and CD1b cells were demonstrated by immunohistochemistry, and immunoglobulin-containing cells (ICC) of the IgA, IgG and IgM classes were demonstrated by immunofluorescence. The labeled cells present in the mucosa of both organs were counted with an image analyzer. The total number of cells was compared between both tissues and differentially between the epithelium and the connective tissue of the mucosa. Significant differences were found in the total number of CD4, CD45RO, and WC1 lymphocytes, in CD14 macrophages, and CD1b dendritic cells, with mean values being greater in the fornix than in the urethra (p<0.05) in all cases. Only dendritic cells were found in the prepuce. No differences were found in the number of CD8 lymphocytes between both organs. The ratio between each cell type in the connective and the intraepithelial tissues and between organs was 10/1 for CD4 in the fornix (p<0.05), against 7/1 in the urethra (p<0.05), while CD8 had a 1/1 distribution in both mucosae. The WC1 ratio was 5/1 in both mucosae (p<0.05). CD45RO labeling was 19/1 in the prepuce (p<0.05) and 1/1 in the urethra. IgA-containing cells did not show differences in the total number of cells in both tissues. In the urethra, no IgG-containing cells were observed and IgM-containing cells were scarce; in contrast, both cell types were present in the prepuce, in amounts greater than in the urethra (p<0.05). IgA-, IgG-, and IgM-containing cells were located in both organs in the mucosal connective tissue. The presence of antigen-presenting cells, macrophages, and dendritic cells, as well as of lymphocytes CD4, CD8 TCR γδ (WC1), IgA-, IgG and IgM positive cells, and CD45RO cells suggests that both mucosae may behave as inductive and effector sites for the mucosal immune response.

Background : The venom of Centruroides limpidus limpidus (Cll) is a mixture of pharmacologically active principles. The most important of these are toxic proteins that interact both selectively and specifically with different cellular targets such as ion channels. Recently, anticancer properties of the venom from other scorpion species have been described. Studies in vitro have shown that scorpion venom induces cell death, inhibits proliferation and triggers the apoptotic pathway in different cancer cell lines. Herein, after treating human cervical adenocarcinoma (HeLa) cells with Cll crude venom, their cytotoxic activity and apoptosis induction were assessed. Results : Cll crude venom induced cell death in normal macrophages in a dose-dependent manner. However, through viability assays, HeLa cells showed high survival rates after exposure to Cll venom. Also, Cll venom did not induce apoptosis after performing ethidium bromide/acridine orange assays, nor was there any evidence of chromatin condensation or DNA fragmentation. Conclusions : Crude Cll venom exposure was not detrimental to HeLa cell cultures. This may be partially attributable to the absence of specific HeLa cell membrane targets for molecules present in the venom of Centruroides limpidus limpidus. Although these results might discourage additional studies exploring the potential of Cll venom to treat human papilloma cervical cancer, further research is required to explore positive effects of crude Cll venom on other cancer cell lines.

In this work a comprehensive review of ewe pregnancy toxemia is presented. Different thematic sections were considered, for a better understanding of the predisposing factors involved, their pathophysiology, clinical signs, post mortem findings, and the diagnosis. Finally, an analysis of current therapy of the disease is presented.

En este trabajo se presenta una exhaustiva revisión de la toxemia de la gestación en la oveja. La misma ha sido dividida en diferentes secciones para lograr una mejor comprensión de los factores que predisponen a esta enfermedad, su patogenia, signos clínicos, los hallazgos postmortem y el diagnóstico. Por último se presenta un análisis de los distintos tratamientos que se utilizan en esta enfermedad.