Two studies were done to study detoxification of aflatoxin (AF)-contaminated chick feed with Nocardia corynebacteroides (NC). In the first study, pathogenicity of the bacteria was studied; in the second, the nutritional value of detoxified feed was evaluated. Commercial corn was divided into 2 sublots, one of which was contaminated with AF. Both lots were divided into 2 parts; the first was inoculated with NC. Four corn-soybean diets were prepared from the 4 corn lots. A completely randomized design was used with 2 x 2 factorial arrangement in which the factors were AF contaminated or not and NC inoculated or not. One hundred Ross 308 chicks (1-d-old, male) were used in 4 treatments with 5 repetitions and 5 chickens per cage. Bird weight and feed consumption were recorded weekly. Each week, 1 chick per treatment repetition was killed for histopathologic analysis of liver, kidney, bursa of Fabricius, pancreas, and small intestine (duodenum, jejunum, and ileum) and for analysis by scanning electron microscopy of the 3 sections of the intestine. At 21 d (the end of both experiments), 1 chick per treatment repetition was killed, and moisture, lipid content, and residual AF in liver were detected. Results at 3 wk did not show differences between treatments (P > 0.05) in any of the variables. In the second study, the same methodology was used except that greater levels of AF were used (800 and 1,200 mug of AFB1/kg of feed). Results showed differences (P < 0.05) in body weight, lipid content, and residual AF in liver. Histopathologic studies showed statistical differences in lesion severity in liver, duodenum, and kidney. Scanning electron microscopy analysis showed severe lesions of intestinal mucosa that mainly affected tight junctions in AF treatments. It can be concluded that NC is safe for chicks and may be used to partly detoxify chicken feed contaminated with AF.
Mastitis in goats is mainly caused by coagulase-negative Staphylococcus (CNS). The identification methods for this group are based on evaluating the expression of phenotypic characteristics such as the ability to metabolize various substrates; however, this is disadvantageous as these methods are dependent on gene expression. In recent years, genotyping methods such as the Multiple Locus Variable-Number Tandem Repeat Analysis (MLVA) and gene identification have been useful for epidemiological study of several bacterial species. To develop a genotyping method, the genome sequence of Staphylococcus chromogenes MU970 was analysed. The analysis showed nine virulence genes described in Staphylococcus aureus. The MLVA was developed using four loci identified in the genome of S. chromogenes MU970. This genotyping method was examined in 23 strains of CNS isolated from goat mastitis. The rate of discrimination for MLVA was 0.8893, and the highest rates of discrimination per the index of Simpson and Hunter-Gaston were 0.926 and 0.968 for the locus 346_06, respectively. The virulence genes were present in all strains of S. chromogenes but not in other CNS. The genotyping method presented in this paper is a viable and easy method for typifying CNS isolates from mastitis cases in different regions and is an ideal mean of tracking this disease.
The aim of this work was to evaluate the relative gene expression levels of the cytokines IL-1B, IL-8, IL-12, IFN-γ, IL-4, IL-10 and TGF-β in somatic milk cells of French Alpine breed, anestrous goats that were experimentally infected in the left mammary gland with Staphylococcus chromogenes during the lactation peak. Milk samples were obtained from both glands for 21 consecutive days post infection. Total RNA was extracted, and real-time PCR was conducted using primers specific to each cytokine. The relative RNA expression of the evaluated cytokines was determined by the comparative method 2 -ΔΔCT , using milk from the right gland of the goats as a reference (control) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as an endogenous control. According to the Wilcoxon test results, IL-1B and IL-12 expression levels showed significant differences compared to those in the control group (p<0.05) from 24 hours post infection until the end of lactation; on day three, IL1β, IL8, IL12 and TGF-β had a statistically significant change in expression with respect to those in the control group (p<0.05); closer to the end of the lactation period, there is no overexpression of the anti-inflammatory interleukins (IL-4 and TGF-β) which may reflect the effort of the host immune system to eradicate the microorganism from the mammary gland.
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