Risk factors associated with the occurrence of multiresistant Staphylococcus spp. isolated from bovine subclinical mastitis in northern Brazil

Staphylococcus bacteria are often associated with subclinical bovine mastitis. This study aimed to identify multiresistant Staphylococcus spp. associated with subclinical mastitis and the associated risk factors. Twenty-three dairy farms with a history of decrease in milk production, located in the lower Acre region, Brazil, were selected. An epidemiological questionnaire was provided in all farms. All animals were examined using the California Mastitis Test (CMT) and their milk samples were collected for bacterial culture. After isolation and identification, the disk diffusion antimicrobial susceptibility test was performed against nine classes of antimicrobials. Of the 339 cows examined using the CMT, 108 had mastitis. A total of 229 milk samples were collected from individual teats. MALDI-TOF MS found isolates belonging to eight species of Staphylococcus, in 101 of these samples. S. chromogenes (58.4%) demonstrated strongest resistance to the nine classes of antimicrobial active principles. Nineteen isolates with multidrug resistance phenotypic profile were identified. This phenotypic expression indicates wide circulation of resistant genes in this species. The presence of multidrug resistance in Staphylococcus spp. in this study was correlated with lack of water for cleaning the corral, which is a preventive factor, minimizing the transmission and persistence of pathogens in the farms.

​Determination of Serogroup and Lytic Activities of Bacteriophages Isolated from Phage Plaques in Staphylococcus aureus Cultures Identified from Sheep Milk with Mastitis

Backgorund: Bacteriophages are closely related to the evolution and virulence of some important bacterial pathogens. Due to their highly significant roles in pathogenesis and virulence, S. aureus bacteriophages are frequently studied. Bacteriophages are grouped into two main categories depending on their life cycles. There are highly consistently lytic phages (virulent) and temperate phages. This study aimed to isolate bacteriophages and determine their phage serogroups from phage plaques in S. aureus cultures in order to show if they are lytic or lysogenic, the latter plays a major role in horizontal gene transfer. Methods: A total of 234 S. aureus isolates were recovered from milk samples from cases with gangrenous mastitis in sheep. Staphylococcal phages are determined based on the type and serogroup by PCR using specific primers. Result: Our study allowed us to determine serogroups of the isolated bacteriophages. Two phage stock samples included only one serogroup while the others included more than one phage serotypes and needed further purification Fa, L and D serogroups were not determined in the study. Present work revealed that all the isolated phages were temperate phages, which play a highly significant role in horizontal gene transfer.

Correlations between the Total Antioxidant Activity and Biochemical Parameters of Cow Milk Depending on the Number of Somatic Cells

The aim of the work was to study the correlations between the total amount of water-soluble antioxidants (TAWSA) and biochemical parameters (BC) of cow milk depending on the somatic cell count (SCC). The BC and TAWSA values of cow milk were measured by spectroscopic and amperometric methods, respectively. The milk samples from the black-and-white cows (Moscow region) were divided according to SCС values: (1) ≤200, (2) 200-499, (3) 500-999, and (4) ≥1000 thousand units/mL. The average TAWSA values for groups 1, 2, 3, and 4 (33, 15, 13, and 12 milk samples) were the following: 15.95 ± 0.74 , 14.45 ± 0.84 , 16.04 ± 0.63 , and 14.58 ± 1.18 . The correlations between TAWSA and BC (group 1) were the following: total fat percentage (TFP) -0.305; true protein percentage (TP1) -0.197; total nitrogen percentage (TN2) -0.210; lactose -0.156; solids-not-fat (SNF) -0.276; total dry matter (TDM) -0.399; freezing point (FP) -0.112; pH -0.114; somatic cell count (SCC) - (-0,052). The correlations between TAWSA and BC (group 2) were the following: TFP -0.332; TP1 -0.296; TN2 -0.303; lactose - (-0.308); SNF -0.159; TDM -0.391; FP -0.226; pH - (-0.211); SCC -0.193. The correlations between TAWSA and BC (group 3) were the following: TFP - (-0.352); TP1 - (-0.411); TN2 – (-0.401); lactose - (-0.166); SNF - (-0.462); TDM - (-0.504); FP - (-0.766); pH - (-0.047); SCC - (-0.698). The correlations between TAWSA and BC (group 4) were the following: TFP -0.159; TP1 -0.046; TN2 – 0.077; lactose - (-0.317); SNF - (-0.237); TDM -0.058; FP - (-0.036); pH - (-0.477); SCC - (-0.072). These data are important in assessing the physiological-biochemical status and state of the antioxidant defense system of cows’ organism.

STUDIES OF NOCARDIOSIS IN CATTLE 1 - ISOLATION OF NORCARIDA ASTEROIDES FROML MASTITIS IN CATTLE

A six months survey (2-8/1994) was canied out to determine norcaiidal mastitis in cattle in certain dairy herds. A total 126 milk samples collected from mastitic‘ cows and milk tank. Of which 10 from Al-Khalis station, 15 from A1-Wahda station, 6 from White Good village, 90 from Al-Dejaila station and 5 fromMilktauk. Examination of the samples result in isolation of Norcaitlia asteroides from Al-Dejaila station. The results explained that 25 animals were positive for norcaridal infection with prevalence of 20.8 % of these 25 cases, 5 cases, in pure cultural and 20 cases, mixed with other organisms. Bacteriological examination of mill: tank also resulted in positive for norcaridial isolates. Metastasis to the lung and supramammary lymph node was demonstrated. The results also explained that norcardial isolates from mastitic cows were more virulent to guinea pigs, andthis study revealed that norcardial infection are not rare in cattle.

A rapid phage assay for detection of viable Mycobacterium avium subsp. paratuberculosis in milk

Paratuberculosis is an incurable gastroenteritis among ruminants that is promoted by Mycobacterium avium subsp. paratuberculosis (MAP), an acid-fast mycobacterium. To accelerate the detection of viable pathogen, a conventional (peptide mediated magnetic separation: PMS) and novel (phage-bead qPCR: PBQ) phage based assay was optimized. A superior limit of detection (LOD) of 10 MAP per 10 mL milk was suggested for PBQ compared to 100 cells/10 mL for PMS-phage assay. Via PBQ, viable MAP was found in 48.78% out 41 unpasteurized sheep and goat milk samples. Sheep milk samples (n = 29) that were tested by PMS-phage assay contained no viable MAP. The absence of viable MAP in milk collected from 21 of the recent sheep animals was also confirmed by PBQ after a 2-week gap. Although, the two phage assays comparably detected no viable MAP in the milk samples, MAP DNA and antibodies against MAP were recognized in milk and sera of some of these animals within two instances of sampling representing that some sheep animals were MAP shedders. In conclusion, PBQ and PMS-phage could be promising methods for the assessment of MAP viability in milk samples. However, PBQ was privileged over the PMS-phage assay due to the lower LOD, rapidity, higher sensitivity, lack of need to M. smegmatis and consequent virucidal treatment that are essential in PMS-phage assay for making lawn and inactivation of exogenous mycobacteriophages respectively.

Classification of dairy properties according to the geometric mean of the somatic cell count and its antimicrobial resistance profile

Milk and its derivatives are highly consumed foods worldwide, with recognized nutritional importance. The search for the production of products with superior quality is constant. For the present work, 26 milk-producing properties were selected, with a total of 506 milk samples collected during the period from October 2019 to May 2020 being evaluated. The objective of this study was to evaluate the quality of milk produced in dairy properties in the region west Paraná, classified as good or bad based on the results of the Somatic Cell Count (SCC) and through sampling (n = 10) to evaluate the resistance profile of enterobacteria and Staphylococcus spp. isolated from milk samples, in addition to the presence of the mecA gene in strains of Staphylococcus spp. resistant to oxacillin. There were significant differences between the good and bad properties for the levels of lactose, SCC (cell/mL), and Standard Plate Count (SPC) (CFU/mL). The strains of Staphylococcus spp. showed differences in the percentage of resistance in relation to the good and bad properties for antibiotics: tetracycline, ciprofloxacin, oxacillin, amikacin, clindamycin, gentamycin, and erythromycin. The mecA gene was not detected in any of the coagulase-negative Staphylococcus isolates that showed resistance to oxacillin. For enterobacteria, the isolated species differed in relation to the classification of properties, with predominance for Escherichia coli (40%) for properties classified as bad and Hafnia alvei (40%) for those classified as good. The percentage of antibiotic resistance compared to enterobacteria isolates was higher in properties classified as good. Monitoring through microbial culture and antibiogram is extremely important, favoring the correct choice for the treatment of animals with a reduced selection of resistant strains.

Cumulative Dietary Risk Assessment of Benzophenone-Type Photoinitiators from Packaged Foodstuffs

Photoinitiators used in ultraviolet-cured ink may migrate from food packaging materials into food products. Therefore, we conducted a dietary risk assessment of exposure to benzophenone (BP)-type photoinitiators by quantifying and reducing uncertainties associated with the risk characterization. A total of 362 food packaging samples including 180 cereals, 136 fruit and vegetable juices, and 46 milk samples were subjected to fast pesticides extraction to determine photoinitiator residues. The average daily dose (ADD) of BP was the highest in the age group of zero to three years, with a P97.5 ADD of 2.56 × 10−4 mg/kg bw/day. The ADD of 2-hydroxybenzophenone (2-OHBP) was the highest in the age group of three to six years, with a P97.5 UB ADD of 3.52 × 10−5 mg/kg bw/day. The estimated UB P97.5 ADD for each age group was below the toxicological concern threshold of 0.0015 mg/kg bw/day. The cumulative toxicity of all BPs, evaluated using the MOET value, was at an acceptable level. Although the MOET value of BPs was above the safety limit in the foodstuffs studied herein, this result may be different if Taiwan were to follow regulation guidelines for BP-type photoinitiators based on the specific migration limit for the unmeasured BP residues in other foodstuffs.

Baseline composition and microbial quality assessment of raw milk from endemic small ruminants and Maghrebi camels in the oasis area of Tunisia

Characteristics and quality aspects of milk from native ovine queue fine de l’Ouest (QFO) and the local goat population were investigated and compared with those of the local Maghrebi camel. A total of 378 individual milk samples were collected from lactating animals reared in the continental oasis region of Tunisia. Samples were analyzed for physical parameters (pH, density, and acidity), chemical composition (dry matter, fat, protein, lactose, casein, ash, and casein-protein ratio), mineral concentrations (Ca, P, Na, and K) and microbiological features (total mesophilic aerobic bacteria (TMAB), total coliform count (TCC), lactic acid bacteria (LAB), sulphite-reducing Clostridium (CSR), yeast and molds (Y/M), Staphylococcus aureus (S. aureus), Escherichia coli (E. coli), and Salmonella) according to standard methods. The results obtained for milk characteristics revealed noticeable disparities between the three species. The mean values of pH, density, and acidity in milk collected from sheep appeared higher than those in milk collected from goat species. Compared to the camel populations, sheep species produced milk with similar pH but higher density and acidity. Between camel and goat species, pH and acidity were higher in Negga, while the density was similar. For milk composition, the results showed a remarkable variation among all studied species and an obvious superiority of the ovine species over the caprine and camel populations in all the chemical contents being studied, except for the casein-protein ratio, which is in favor of goat species. The milk of QFO sheep, the richest in casein and protein, was expressed with significantly higher levels of calcium and phosphorus than goat and camel milk. Compared to small ruminants, milk from camels is the richest in Na and K. Additionally, more Ca is present in milk from camels than goats. Goat milk, the poorest type of milk in Ca and Na, contains on average more P than camel milk and more K than sheep's milk. The poor bacteriological quality was that of camel milk for all microbial counts. The microbial quality of goat milk was higher than that of ewe milk based on TMAB, TCC, and E. coli counts, while ovine milk was of better quality, referring to LAB, Y/M, and S. aureus values. No significant differences were found for Staphylococcus aureus and Escherichia coli between the examined species. The obtained results highlighted the complete absence of the two dangerous pathogens Salmonella and CSR in all investigated milk samples. The microbiological examination evidenced that the milk of small ruminant species complies with standard criteria required by Tunisian legislation on the hygiene of milk and dairy products. Regarding camel milk, the microbial analysis revealed poor quality that exceeds standard criteria.