Analysis of Escherichia coli isolated from bovine mastitic milk

Mastitis has been recognized for some time as the most costly disease in dairy herds. From February to November 2004, 670 samples of bovine mastitic milk from which 231 Escherichia coli strains were isolated, were collected from two Brazilian states. The strains were screened for the presence of Shiga toxin-producing (stx 1 and stx 2) and intimin (eae) genes. Twenty (8.6%) strains were detected by PCR to harbor the Shiga toxin genes (8 the stx 1 gene, 12 the stx 2 gene and none both of them). Two (0.8%) of the Escherichia coli strains studied were eae positive non Shiga toxin-producing. The strains were also examined for resistance to 12 antimicrobial agents. The predominantly observed resistance was to tetracycline (92.2%), streptomycin (90.4%), nalidixic acid (88.3%), amikacin (86.5%) and cephalothin (84.8%). Multidrug resistance was found among 152 isolates (65.8%).

Morphological aspects of buffaloes (Bubalus bubalis) umbilical cord

Buffalo is an important livestock resource, with a great participation in agricultural systems, providing milk, meat, and work power. Umbilical cord is responsible for maternal-fetal nutrients exchange during pregnancy, and its alterations can compromise the fetal development. We investigated ten pregnant uteruses collected from cross-bread buffaloes in different stages of gestation. Pregnancy and fetal age was determined by measuring the apex sacral length and development period was calculated by previously published formula. Umbilical cords were measured for length determination. Umbilical cord vascular net and anastomosis were observed by injection of Neoprene latex. Histological sections of the umbilical cord were studied after stain with HE, picrossirius, toluidine blue, orceine, and PAS reaction. Buffaloes' umbilical cord was formed by two central arteries, an allantois duct and two peripheral veins. The artery wall was composed by large quantity of collagen, elastic fibers, fibroblasts and large number of vasa vasorum. The allantois duct was located between the arteries and presented a great number of small nourishing vessels. Small nourishing vessels should be carefully considered to avoid to be mistaken to the arterials and veins vasa vasorum. Medium length of umbilical cord from buffalos was 11.8cm (minimum of 6.8cm and maximum of 17.4cm).

Organophosphorus and carbamates residues in milk and feedstuff supplied to dairy cattle

Considering acute and chronic toxicity effects on human and animal health caused by pesticide residues in food, this study aimed to analyze organophosphorate (OP) and carbamate (CB) in feedstuff and water destined for dairy cattle, as well as in the milk produced by these animals, through gas chromatography (GC). In the Agreste region of Pernambuco, Brazil, 30 raw milk samples and all components of the animals' diet were collected from several farms. Out of the 30 milk of milk analyzed, six (20%) were contaminated with OP, five (16.7%) with CB, and one sample with both pesticides. From 48 analyzed feed samples, 15 (31.25%) were contaminated with residues of OP, six (12.50%) with CB, and one sample was contaminated with both pesticides. Out of 16 water samples analyzed, six (37.50%) were contaminated with OP residues, but non with CB. In four dairy farms the pesticides detected in milk were compatible with the active principles found in water and/or foodstuff, suggesting them to be the source of contamination.

Breed and season influence on milk quality parameters and in mastitis occurrence

The aims of the present study were to evaluate the performance of Jersey and Holstein cows under different rainfall conditions (dry and rainy seasons) by monitoring aspects related to subclinical mastitis (somatic cell count, microbiological isolation, type of isolated pathogen), milk quality (lactose, protein, fat, total solids) and production (mean milk production) of both breeds. The study was carried out in a dairy farm located in the state of São Paulo, Brazil. Eight visitations were done to the farm, four in a period of high rainfall and four in a period of low rainfall. Milk samples were collected from 79 Holstein cows and 37 Jersey cows for electronic somatic cell count and determination of the main milk components (protein, fat, total solids, lactose). Milk fat, protein, total solids and production were influenced by breed and the season, with similar tendencies for both breeds in both seasons. Somatic cell count (SCC) showed similar results for both breeds. Holstein cows with intramammary infections (IMI) presented a higher increase in SCC when compared to Jersey cows (P<0.001). In the dry season, 53 animals had IMI in at least one month during the study, which 32 were Holstein and 21 were Jersey cows. In the rainy season, 65 animals had intramammary infection, being 43 Holstein and 22 Jersey cows. The frequency of IMI cases was larger in the rainy season than in the dry season. Jersey cows had a lower chance of showing IMI signs and symptoms than Holstein cows in the rainy season (odds ratio=0.52). The larger number of IMI cases in the rainy season may have led to a lower milk lactose rate for both breeds, thus milk lactose rate can be considered an indicator of IMI status. There was prevalence of contagious pathogens overall in the study. The applied model showed that environmental pathogens were more frequently isolated from the breed Jersey, regardless of the study season. There seems to be differences in the immune response of Jersey and Holstein breeds.

Sanitizer residues in the milk of cows which had passed through footbaths

Copper sulfate and sodium hypochlorite are used in footbath solutions for the prevention and treatment of bovine digital diseases; however, data on the residues of such elements in milk are sparse in Brazil. This study evaluated the cost of applying the footbath treatment and the total amount of copper and chlorite residues in the milk of healthy cows after they had passed through these footbath solutions. Two groups of 7 cows each (GI and GII) were studied. In the case of GI, 1% sodium hypochlorite was used and for GII 5% copper sulfate was employed in the footbath. The milk samples were collected before the 7-day footbath treatment period (M0) and 24 h (M1), 48 h (M2), 72 h (M3) and 15 days (M15) after the last footbath. Statistical analysis to compare the different samples within each group was carried out by applying Friedman's test, followed by Dunn's test (p<0.05). It was concluded that the amount of total chlorites and copper in the milk of healthy cattle after routine daily footbaths for a period of 7 days presented some variations. However, the concentrations observed were considered insufficient to represent a risk to human health. The cost of the footbath solutions was found to be reasonable.

Mycoplasma agalactiae in semen and milk of goat from Pernambuco state, Brazil

In goat and sheep flocks, mycoplasmosis is a disease that may cause severe economical losses associated with polyarthritis, mastitis, agalactia, conjunctivitis, pneumonia and reproductive failure. The latter may involve repeat breeding, granular vulvovaginitis, infertility and abortions. The aim of the present study was to assess the occurrence of Mycoplasma agalactiae (Ma) in semen and milk samples from naturally infected goat in the semiarid region from Pernambuco State, Northeast from Brazil. Thirty-nine semen samples and 81 milk samples were submitted to DNA extraction using a commercially available kit and following the manufacturer's instructions. The polymerase chain reaction (PCR) was then performed in accordance with protocols described in the literature. The results of the present study revealed the presence of Ma in the DNA of 17.9% (7/39) of the semen samples and 3.7% (3/81) of the milk samples. The results obtained in the present study confirm the elimination of the DNA of Ma in the semen and milk samples. The presence of this agent in goat flocks is considered very risky in terms of reproductive disorders and contagious agalactia outbreaks in the Northeast region of Brazil.

Acid-base balance of dairy cows and its relationship with alcoholic stability and mineral composition of milk

This study aimed to associate the occurrence of acid-base disorders with the alcoholic stability of milk from animals in the field, and to evaluate differences between the mineral composition of milk that was both stable and unstable in alcohol. The sample comprised 96 dairy cows, where the milk and blood of each corresponding animal was collected. The mineral composition of stable and unstable milk in alcohol was different and may be related to acid-base disturbances. The average amount of phosphate was lower in the milk that was unstable in alcohol, while potassium was greater. Frequency of the alcoholically unstable milk cases was higher in the cows with acid-base disturbances. Respiratory alkalosis was the disorder that was most observed.

Elimination of the tremorgenic toxin of Ipomoea asarifolia by milk

With the aim to determine if the tremorgenic toxin of Ipomoea asarifolia is eliminated in milk, three groups of Swiss female mice received, immediately after giving birth until weaning, a ration containing 20% or 30% of dry I. asarifolia. All the offspring of the females that received 20% or 30% I. asarifolia showed tremors 2-4 days after birth. The offspring of the females that received 20% I. asarifolia recovered 4-7 days after weaning. The offspring of the females that received 30% of the plant in the ration died while showing tremors before weaning or up to two days after weaning. It is concluded that the tremorgenic compound of I. asarifolia or its toxic metabolites are eliminated in milk, and that lactating mice may be used as a model for the determination of the toxic compound(s) in this plant.

Expression of CD14 and toll-like receptors 2 and 4 by milk neutrophils in bovine mammary glands infected with Corynebacterium bovis

This study evaluated the expression of CD14, toll-like receptor (TLR) 2 and TLR4 on the surface of milk neutrophils in bovine mammary glands infected with Corynebacterium bovis. Here, we used 23 culture-negative control quarters with no abnormal secretion on the strip cup test and milk somatic cell count lower than 1x105 cells/mL, and 14 C. bovis infected quarters. The identification of neutrophils, as well as, the percentage of neutrophils that expressed CD14, TLR2 and TLR4 were analyzed by flow cytometry using monoclonal antibodies. The present study encountered no significant difference in the percentages of milk neutrophils that expressed TLR2 and TLR4 or in the expression of TLR4 by milk neutrophils. Conversely, a lower median fluorescence intensity of TLR2 in milk neutrophils was observed in C. bovis-infected quarters. The percentage of neutrophils that expressed CD14 and the median fluorescence intensity of CD14 in milk neutrophils was also lower in C. bovis-infected quarters.

Partial budget analysis of prepartum antimicrobial therapy and Escherichia coli J5 vaccination of dairy heifers and their effect on milk production and milk quality parameters

Abstract: This study aimed to determine whether prepartum antimicrobial and/or Escherichia coli J5 vaccination in dairy heifers influence the milk production, milk quality, and estimate their economic benefit. Thus, 33 dairy heifers were enrolled in four groups using a split-splot design. Groups were: (G1) prepartum antimicrobial infusion and vaccination with an E. coli J5 bacterin, (G2) prepartum antimicrobial infusion, (G3) vaccination with an E. coli J5 bacterin, and (G4) control heifers. Composite milk samples for somatic cell count, total bacteria count and milk composition were collected 15 days after calving and every 15 days until the end of the experiment. Bacteriological analysis was carried out at the end of study. The milk production and the incidence of clinical cases of mastitis, as well as the costs associated with them were recorded. The results demonstrate a reduction on clinical mastitis rates by preventive strategies, which implicated in lower volume of discarded milk (0.99, 1.01, 1.04 and 3.98% for G1, G2, G3 and G4, respectively) and higher economic benefit. Thus, in well-managed dairy herds the prevention of heifer mastitis by vaccination or antimicrobial therapy can reduce the amount of antimicrobials needed to treat clinical mastitis cases and the days of discarded milk.

Milk protein polymorphisms in Brazilian Zebu cattle

Five bovine milk protein polymorphisms were studied in Zebuine cattle raised in Brazil, through horizontal electrophoresis on starch gel containing urea and 2-mercaptoethanol, using basic and acidic buffer systems. Allelic frequencies for a-La, b-Lg, aS1-Cn, b-Cn and k-Cn loci were estimated in six Gyr herds (N = 283), six Guzerat herds (N = 205), one Nelore herd (N = 17) and one Sindi herd (N = 22), all from São Paulo or Minas Gerais State, Brazil. Genotypic frequencies observed for each locus and breed studied are in accordance with the assumption of genetic equilibrium, demonstrating absence of high inbreeding levels for the breeds tested. The FST value found indicated significant genetic differentiation among breeds; however, the Gyr and Guzerat herds showed significantly different gene frequencies. Genetic distance estimates among zebuine breeds studied and the Holstein breed, taken as a reference for a taurine breed, showed strong differences between these two racial groups

Morphological changes of the jejunal mucosa in protracted diarrhea and their correlation with disease duration, weight loss and serum albumin levels

The pathogenesis of protracted diarrhea is multifactorial. In developing countries, intestinal infectious processes seem to play an important role in triggering the syndrome. Thirty-four children aged 1 to 14 months, mean 6.5 months, with protracted diarrhea were studied clinically and in terms of small intestinal mucosal morphology. Mild, moderate or severe hypotrophy of the jejunal mucosa was detected in 82% of cases, and mucosal atrophy was observed in 12%. The intensity of the morphological changes of the jejunal mucosa correlated negatively with serum albumin levels. No correlation was detected between mucosal grading and duration of diarrhea or between mucosal grading and weight reported as percentile. After nutritional support was instituted, serial jejunal biopsies were obtained from 12 patients: five patients submitted to parenteral nutrition for 7 to 38 days, mean 17 days, and 7 patients receiving a hypoallergenic oral diet (semi-elemental formula, 3; chicken formula, 3; human milk, 1). In seven cases (58%) a progressive increase in villus height and a decrease in the number of inflammatory cells were noted. Recovery of the morphologic pattern was accompanied by clinical improvement in all patients

Effects of ethanol intake on retinol concentration in the milk of lactating rats

The effect of the consumption of ethanol (5%) on retinol concentration in milk was studied in the rat on day 12 after delivery, together with the evolution of dam body weight and pup growth rate. Female Wistar rats receiving alcohol (5%) in drinking water during lactation (N = 7) were compared to normal controls fed ad libitum (N = 6). The mean maternal alcohol intake was 3.96 ± 0.23 g/kg body weight per day. To determine retinol levels in milk we used the Bessey and Lowry method, modified by Araújo and Flores ((1978) Clinical Chemistry, 24: 386-392). The pups were separated from dams for a 2-4-h period, after which the dams were injected intraperitoneally with anesthetic and oxytocin. The concentration of retinol in milk was 162.88 ± 10.60 µg/dl in the control group and 60.02 ± 8.22 µg/dl in the ethanol group (P<0.05). The ethanol group consumed less food than the controls and lost a significant amount of weight during lactation. On days 8, 10 and 12, the body weight of the pups from rats given ethanol (13.46 ± 0.43, 16.12 ± 0.48 and 18.60 ± 0.91 g, respectively) were significantly lower (P<0.05) than the weight of pups from controls (15.2 ± 0.44, 18.36 ± 0.54, 20.77 ± 0.81 g). These data show that ethanol intake during the suckling period, even at low concentrations, decreases the amount of retinol in milk and, therefore, the amount available to the pups.

BCG lymphadenopathy detected in a BCG-vaccinated infant

Large-scale vaccination with BCG, the live attenuated strain of Mycobacterium bovis, is being adopted around the world, although sporadic complications have occurred after the procedure. Lymphadenopathy is not uncommon especially in babies under one year (0.73% of vaccinated infants), but the swelling subsides within 2 months in most cases, with no medical or surgical treatment. Brazil adopted BCG vaccination program earlier in the seventies and by 1995 more than 96% of the infant population received this immunization. We report here the occurrence of lymphadenopathy in a two-year-old child vaccinated with the Brazilian BCG strain. The diagnosis was made using a lymph node biopsy and intestinal aspirates that yielded a positive mycobacterial culture. The isolate was resistant to isoniazid, rifampicin, pyrazinamide and thiophen-2-carbonic acid hydrazide, sensitive to streptomycin, ethambutol, and p-nitrobenzoic acid, and reacted positively to cyclo-serine and negatively to niacin. The pncA gene involved in bacterial activation of pyrazinamide contains in M. bovis a point mutation that renders pyrazinamidase unable to catalyze drug activation. Therefore, this polymorphism is a good option for developing methods to differentiate M. bovis and M. tuberculosis. Taking advantage of this difference we further analyzed the isolates by single-stranded conformation polymorphism electrophoresis of DNA following PCR of the pncA gene. The isolate identity was confirmed by RFLP electrophoretic analysis of the amplified fragment following Eco065I digestion, which selectively cleaves M. tuberculosis DNA. From this result it is proposed that RFLP of pncA gene represents an alternative for differential diagnosis of M. bovis.

Risk factors for excess weight loss and hypernatremia in exclusively breast-fed infants

Data were prospectively obtained from exclusively breast-fed healthy term neonates at birth and from healthy mothers with no obstetric complication to determine risk factors for excess weight loss and hypernatremia in exclusively breast-fed infants. Thirty-four neonates with a weight loss > or = 10% were diagnosed between April 2001 and January 2005. Six of 18 infants who were eligible for the study had hypernatremia. Breast conditions associated with breast-feeding difficulties (P < 0.05), primiparity (P < 0.005), less than four stools (P < 0.001), pink diaper (P < 0.001), delay at initiation of first breast giving (P < 0.01), birth by cesarean section (P < 0.05), extra heater usage (P < 0.005), extra heater usage among mothers who had appropriate conditions associated with breast-feeding (P < 0.001), mean weight loss in neonates with pink diaper (P < 0.05), mean uric acid concentration in neonates with pink diaper (P < 0.0001), fever in hypernatremic neonates (P < 0.02), and the correlation of weight loss with both serum sodium and uric acid concentrations (P < 0.02) were determined. Excessive weight loss occurs in exclusively breast-fed infants and can be complicated by hypernatremia and other morbidities. Prompt initiation of breast-feeding after delivery and prompt intervention if problems occur with breast-feeding, in particular poor breast attachment, breast engorgement, delayed breast milk "coming in", and nipple problems will help promote successful breast-feeding. Careful follow-up of breast-feeding dyads after discharge from hospital, especially regarding infant weight, is important to help detect inadequate breast-feeding. Environmental factors such as heaters may exacerbate infant dehydration.