Effect of Milk Fermented with Lactobacillus acidophilus NCDC15 on Nutrient Digestibility, Faecal Biomarkers and Immune Response in Murrah calves

Abstract In neonates, rapid change in diet imbalances gut health allowing colonization of opportunistic pathogens that confer harmful effects on animal health causing reduced digestion and malabsorption of nutrients. In this milieu, probiotic feeding can be a promising approach in promoting animal health and stabilization of gastrointestinal microbiota. Hence, the present study was designed to investigate the effect of Lactobacillus acidophilus NCDC15 enriched fermented milk on nutrient digestibility, faecal biomarkers and immune response in Murrah buffalo calves. Twenty-four, neonatal calves (5-7 days) were randomly allocated into four groups for 90 days. The control group (CT) was provided a basal diet of calf starter and green fodder (maize and jowar), without any probiotic fermented milk (PFM) supplementation. Basal diet was supplemented with probiotic fermented milk at 100, 200 and 300 mL/calf/day, in PFM100, PFM200 and PFM300 groups, respectively. Nutrient digestibility remained similar among the groups. Faecal acetate was higher (P<0.05) in PFM200 and PFM300, while, faecal butyrate was increased (P<0.05) with all levels of probiotic supplementation than control. Faecal Lactobacillus and Bifidobacterium count were increased (P<0.05) with a concomitant reduction in coliform population (P<0.05) among all the treatments. Cell-mediated and humoral immune response were higher (P<0.001) in PFM200 and PFM300 than CT. Overall, it can be concluded that inclusion of Lactobacillus acidophilus NCDC15 in the form of fermented milk upto 300 mL/calf/day improved immunity and faecal biomarkers in Murrah buffalo calves without any adverse effect on nutrient utilization which may positively impact growth performance in Murrah buffalo calves.

In-vitro Plasma Protein Binding of Marbofloxacin in Healthy and Disease Condition of Buffalo Calves

Marbofloxacin is a broad-spectrum fluoroquinolone antibiotic developed for use in veterinary medicine for the treatment of skin and soft tissue infections in dogs and cats. Plasma protein binding plays a vital role in distribution, elimination and therapeutic effectiveness of drugs. In the present study we evaluated the plasma protein binding of marbofloxacin in healthy and liver dysfunctioned buffalo calves. In vitro binding of marbofloxacin to plasma proteins was determined by employing the equilibrium dialysis technique and further analyzed by High Performance Liquid Chromatography assay. The plasma protein binding for healthy calves ranges between 25.3±0.34% to 30.4±0.40% with an overall binding of 28.66 ± 0.421%. Kinetic constants (βi) and (Kβ) was 2.6±0.12×10-8 mole/g and 1.9±0.08×10-7 mole, respectively. The percentage of plasma protein binding for liver dysfunctioned buffalo calves extended from 24.5 - 30.3% with an overall mean of 28.59 ± 0.693%. The binding capacity of the drug to plasma proteins (βi) and dissociation rate constant of protein drug complex (Kβ) were 2.53±0.13 10-5 mole/g and 1.94±0.09×10-6 mole respectively. There was no significant change observed in plasma protein binding and the kinetic constant of liver dysfunctioned buffalo calves when compared to the healthy group

Virulence factors and phylotyping of Escherichia coli isolated from non-diarrheic and diarrheic water buffalo calves / Caracterização dos fatores de virulência e filogrupos de Escherichia coli isoladas de bezerros bubalinos sem diarreia e diarréicos

ABSTRACT: This study aimed to determine the virulence factors, phylogenetic groups, and the relationships between pathovars and phylogenetic groups of E. coli strains isolated from feces of buffalo calves. A total of 217 E. coli strains were obtained from feces after culture and were screened by PCR for detection of virulence factors EAST-1, enterohemolysin, Saa, CNF2, F41, F5, STa, intimin, Stx1 and Stx2. One hundred and thirty-four isolates were positive for one or more virulence factors: eighty-four from diarrheic animals, and fifty from non-diarrheic calves. The pathovars of E. coli identified in diarrheic feces were ETEC (F5+) (2/84), NTEC (16/84), STEC (20/84), EPEC (3/84), EHEC (3/84), and EAEC (EAST-1+) (33/84). Pathovars identified in non-diarrheic animals were NTEC (21/50), STEC (17/50), EHEC (1/50) and EAEC (7/50). E. coli strains positive for EAST-1 (P=0.008) and phylogroup C (P = 0.05) were associated with the presence of diarrhea. Phylogenetic analysis showed that 58.95% of the isolates belonged to phylogroup B1, followed by E (9.70%), B2 (5.90%), C (5.90%), D (5.22%), A (2.24%), and F (1.50%). Phylogroup B1 predominated in pathogenic E. coli isolated from water buffalo, and phylogroup C constituted an enteropathogenic E. coli for water buffalo calves.

RESUMO: O objetivo foi determinar os fatores de virulência, os grupos filogenéticos e as possíveis relações entre os patovares e os grupos filogenéticos identificados de cepas de Escherichia coli isoladas de fezes de bezerros bubalinos. Um total de 217 amostras de E. coli foram identificadas a partir de cultura das fezes e submetidas a reação em cadeia da polimerase (PCR) para detecção dos fatores de virulência EAST-1, enterohemolisina, Saa, CNF2, F41, F5, STa, intimina, Stx1 e Stx2. Foram identificadas 134 cepas positivas para um ou mais fatores de virulência: 84isoladas de bezerros bubalinos diarreicos e 50 de bezerros bubalinos saudáveis. Os patovares de E. coli obtidos de fezes diarreicas foram ETEC (F5+) (2/84), NTEC (16/84), STEC (20/84), EPEC (3/84), EHEC (3/84), e EAEC (EAST-1+) (33/84). Os patovares isolados de fezes não diarreicas foram NTEC (21/50), STEC (17/50), EHEC (1/50) e EAEC (7/50). Cepas de E. coli positivas para EAST-1 (P = 0,008) e filogrupo C (P = 0,05) foram associadas com a presença de diarreia. A análise de filogrupos revelou que 58,95% dos isolados pertencem ao filogrupo B1, seguido por E (9,70%), B2 (5,90%), C (5,90%), D (5,22%), A (2,24%) e F (1,50%). O filogrupo B1 predomina em cepas de E. coli patogênicas isoladas de bezerros búfalos e o filogrupo C constitui um filogrupo de E. coli patogênica entérica para bezerros.

Lymphoglandular complexes in proximal colon of buffalo calves (Bubalus bubalis) / Complejos linfoglandulares en colon proximal de terneros de buffalo (Bubalus bubalis)

The present study was conducted on six healthy early neonatal and six prepubertal buffalo calves to study the location, gross morphology, histomorphology and histochemistry of lymphoglandular complexes in proximal colon. In very proximal part of colon of buffalo calves, an irregular oval mucosal lymphoid patch was found grossly as a proximal colon (PC) patch. Histologically, in proximal colon patch of early neonates (3-4 weeks), an extensive invasion of mucosal glands was observed towards lymphoid nodules that were present in submucosa. The structure as a whole thus formed a complex known as lymphoglandular complex (LGC). Large number of such complexes i.e., LGCs were observed in submucosa of proximal colon at this age. At some places, invasion of mucosal glands into lymphoid tissue was restricted to superficial layer of complexes, with the lymphoglandular complexes opening directly into the lumen but some were deep seated. However, by the age of 6 months in buffalo calves i.e., prepubertal period, LGCs were reduced and were present in single layer within the submucosa of the proximal colon. Moreover, some of LGCs were completely encapsulated by their own lamina muscularis mucosae. But some of the complexes still had their mucosal openings into lumen while others had lost their connection with tunica mucosa. Histochemically, the glands that were observed within LGCs contained mucosubstances, glycogen, mucopolysaccharides, and mucin. However, lipids were present around the lymphocytes observed towards the periphery of these LGCs.

El presente estudio se llevó a cabo en seis terneros de búfalo neonatos sanos y seis terneros prepuberales para estudiar la ubicación, morfología macroscópica, histomorfología e histoquímica de los complejos linfoglandulares en el colon proximal. Se observó en un área del colon proximal (AP) de los terneros de búfalo un óvalo linfoide de mucosa irregular en la parte más proximal de éste. Histológicamente, en el área proximal del colon de los terneros neonatos (3-4 semanas), se observó una invasión extensa de las glándulas mucosas hacia los nódulos linfáticos presentes en la submucosa. La estructura en su totalidad formaba un complejo conocido como complejo linfoglandular (CLG). A esta edad se observó un gran número de estos complejos es decir, se observaron CLGs en la submucosa del colon proximal. La invasión de las glándulas mucosas en el tejido linfoide, se limita a la capa superficial de los complejos, los complejos linfoglandulares distribuidos directamente en el lumen, sin embargo otros se encontraban arraigados de manera profunda. En búfalo a los 6 meses de edad, es decir en el período prepuberal, se observó un número reducido de CLGs presentes en una sola capa dentro de la submucosa del colon proximal. Por otra parte, algunos de CLGs estaban completamente encapsulados por su propia lamina muscularis mucosae. Algunos de los complejos mantenían abertura de las mucosas en el lumen, mientras que otros habían perdido su conexión con la mucosa. En análisis histoquímico, las glándulas que se observaron dentro del CLGs contenían mucosustancias, glucógeno, mucopolisacáridos y mucina. Sin embargo, se encontraron lípidos presentes alrededor de los linfocitos hacia la periferia de los CLGs.

Plasma levels, pharmacokinetics and dosage regimen of intravenously administered gatifloxacin in buffalo calves (Bubalus bubalis) on coadministration with meloxicam / Os níveis plasmáticos, farmacocinética e regime de dosagem de gatifloxacina administrado por via intravenosa em bezerros búfalos (Bubalus bubalis) na administração concomitante com meloxicam

The pharmacokinetics of intravenously administered gatifloxacin, upon concomitant administration with meloxicam was investigated in buffalo calves. Meloxicam was administered subcutaneously (0.5 mg.kg-1) immediately followed by intravenous administration of Gatifloxacin (4 mg.kg-1). The concentration of gatifloxacin was estimated in plasma by microbiological assay. Pharmacokinetic parameters were calculated and appropriate dosage schedule was computed. The therapeutic plasma drug concentration was maintained up to 12 h. Gatifloxacin was rapidly distributed from blood to tissue compartment, which was evident from the high values of distribution rate constant, α1 (11.9 ± 0.52 h-1) and the ratio of rate constant of transfer of drug from central to peripheral compartments and vice versa, K12/K21 (3.05 ± 0.36) and K13/K31 (2.04 ± 0.12). The area under the plasma drug concentration-time curve and apparent volume of distribution were 12.0 ± 0.68 µg.ml-1.h and 2.69 ± 0.14 L.kg-1, respectively. The elimination half-life (t1/2β), total body clearance (ClB) and the ratio of drug present in peripheral to central compartment (P/C) were 5.59 ± 0.40 h, 337.6 ± 19.9 ml.kg-1.h-1 and 8.04 ± 0.50, respectively. The present study revealed that the most suitable dosage regimen of gatifloxacin when concomitantly administered with meloxicam in buffalo calves would be 2.5 mg.kg-1 followed by 2.0 mg.kg-1 at 12 h intervals.

Investigou-se a farmacocinética da gatifloxacina, administrada por via intravenosa, concomitante à aplicação de meloxicam em bezerros búfalos. O meloxicam foi administrado por via subcutânea (0,5 mg.kg-1), imediatamente seguido pela administração intravenosa de gatifloxacina (4 mg.kg-1). A concentração plasmática de gatifloxacina foi estimada por ensaio microbiológico. Os parâmetros farmacocinéticos foram calculados e a posologia adequada foi computada. A concentração plasmática do fármaco-terapêutico foi mantida por 12 h. A gatifloxacina foi rapidamente distribuída a partir de sangue para o compartimento de tecido, o que ficou evidente a partir dos valores elevados da taxa constante de distribuição, α1 (11.9 ± 0.52 h-1) e a proporção de velocidade constante de transferência de droga a partir de centrais para os compartimentos periféricos e vice-versa, K12/K21 (3.05 ± 0.36) e K13/K31 (2.04 ± 0.12). A área sob a curva plasmática de concentração-tempo da droga e o volume aparente de distribuição foi de 12.0 ± 0.68 µg.ml-1.h e 2.69 ± 0.14 L.kg-1, respectivamente. A meia-vida (t1/2β), a depuração corporal total (ClB) e relação da droga presente no sangue periférico para o compartimento central (P/C) foram 5.59 ± 0.40 h, 337.6 ± 19.9 ml.kg-1.h-1 e 8.04 ± 0.50, respectivamente. O presente estudo revelou que o regime de dosagem mais adequado de gatifloxacina quando administrada concomitantemente com meloxicam em bezerros búfalos seria 2,5 mg.kg-1 seguida de 2,0 mg.kg-1 em intervalos de 12 h.