Enterohemorrhagic Escherichia coli O157: H7 from healthy dairy cattle in Mid-West Brazil: occurrence and molecular characterization

Enterohemorrhagic Escherichia coli (EHEC) serotype O157:H7 represents the major Shiga toxin-producing E. coli (STEC) strain related to large outbreaks and severe diseases such as hemorrhagic colitis (HC) and the potentially lethal hemolytic uremic syndrome (HUS). The aim of this study was to report the occurrence and molecular characterization of O157:H7 isolates obtained by rectal swab from 52 healthy dairy cattle belonging to 21 farms in Mid-West of Brazil. Detection of 16SrRNA, stx1, stx2, rfbO157, fliCh7, eae, ehxA, saa, cnf1, chuA, yjaA and TSPE4.C2 genes was performed by PCR. The isolates were further characterized by serotyping. Two hundred and sixty E. coli isolates were obtained, of which 126 were characterized as STEC. Two isolates from the same cow were identified as serotype O157:H7. Both isolates presented the stx2, eae, ehxA, saa and cnf1 virulence factor genes and the chuA gene in the phylogenetic classification (virulent group D), suggesting that they were clones. The prevalence of O157:H7 was found to be 1.92% (1/52 animals), demonstrating that healthy dairy cattle from farms in the Mid-West of Brazil are an important reservoir for highly pathogenic E. coli O157:H7.

Performance of dairy heifers raised on Xaraés palissadgrass (Brachiaria brizantha cv. Xaraés) pasture supplemented with two types of mineral mixture

The effects of selective (e.g. mineral supplement formulated on the basis of nutritional and clinical examination of the herd) or commercial mineral supplementation of crossbred dairy heifers (Holstein-Mantiqueira), on daily weight gain (DWG), body condition score (BCS), age at first mating (AFM) and the intake of mineral mixture (IMM) managed on Xaraés palissadgrass pasture (Brachiaria brizantha cv. Xaraés) were evaluated from February 2006 to March 2008. Structural characteristics, forage allowance and nutritional value of Xaraés palissadgrass were also evaluated. The structural variables of pasture, allowances and nutritive value of forage, besides the DWG were only affected (p<0.05) by season, with highest results for spring and summer. Throughout the experimental period (730 days), no sign of mineral deficiency was detected in heifers that ingested the selective supplement. The DWG and AFM were not influenced by the mineral mixture offered (0.52 and 0.33 kg/day during the spring/summer and in fall/winter for DWG, respectively, and 813 days of AFM). The IMM was higher for commercial mineral mixture than for selective (61.6 and 51.0g/day respectively). The BCS was slightly higher for selective (3.17) than for the commercial (3.02) mineral mixture. Thus, the selective mineral supplement was a strategy to prevent mineral deficiency in this herd and resulted in similar performance at lower costs (e.g. due to less IMM) of crossbred heifers when compared with the commercial mineral supplementation.

Launching the product innovation to a market: case study from a dairy industry

In the environment of ever-changing needs of customers, technologies and competitors, the survival of the company depends on how well it researches, develops and implements new products to the market. The need for development of new products relates to many factors: globalization, international competition which is now underway on a global scale, scientific advances and development of production, changes in consumer preferences and consumer behavior. In this study the focus is on the company form a dairy products industry. This study is aimed to defining the role of product innovation launch strategy in an overall enterprise strategy, and to select the optimal combination of its marketing tools. The main purpose of this study is to determine place and the role of innovative marketing based in the innovation process, and to determine launch and positioning strategies in the general concept of an innovative product. The object of the study is the Russian enterprise, which is aimed to achieve a competitive advantage through the continuous production of new products, upgrade existing ones and improve innovation management practices. Research showed that the differentiation strategy is suitable for launching the dairy industry product innovation to a market.

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.

Severe diarrhea outbreak in beef calves (Bos indicus) caused by G6P[11], an emergent genotype of bovine rotavirus group A

The episodes of diarrhea caused by neonatal bovine rotavirus group A (BoRVA) constitute one of the major health problems in the calf rearing worldwide. The main G (VP7) and P (VP4) genotypes of BoRVA strains involved in the etiology of diarrhea in calves are G6P[1], G10P[11], G6P[5], and G8P[1]. However, less frequently, other G and P genotypes have been described in BoRVA strains identified in diarrheic fecal samples of calves. This study describes the identification and molecular characterization of an emerging genotype (G6P[11]) in BoRVA strains involved in the etiology of a diarrhea outbreak in beef calves in a cattle herd of high production in extensive management system. The diarrhea outbreak, which showed high morbidity (60%) and lethality (7%) rates, occurred in calves (n= 384) Nelore (Bos indicus) up to 30-day-old from the State of Mato Grosso do Sul, Brazil. BoRVA was identified in 80% (16/20) of the fecal samples analyzed by polyacrylamide gel electrophoresis (PAGE) technique. In all PAGE-positive fecal samples were amplified products with 1,062-bp and 876-bp in the RT-PCR assays for VP7 (G type) and VP4 (VP8*) (P type) of BoRVA, respectively. The nucleotide sequence analysis of VP7 and VP4 genes of four wild-type BoRVA strains showed G6-III P[11]-III genotype/lineage. The G6P[11] genotype has been described in RVA strains of human and animal hosts, however, in calves this genotype was only identified in some cross-sectional studies and not as a single cause of diarrhea outbreaks in calves with high morbidity and lethality rates as described in this study. The monitoring of the G and P genotypes of BoRVA strains involved in diarrhea outbreaks in calves is important for both animal and public health by allowing the identification of the most frequent genotypes, the characterization of novel genotypes and to identify reassortments with genotypes described in animal and human hosts. The results of this study show the importance of the monitoring of the genotypes of BoRVA strains involved in episodes of bovine neonatal diarrhea as for characterization of frequency of occurrence and pathogenic potential of uncommon genotypes as for monitoring of the emergency of different BoRVA genotypes not included in commercial vaccines.

Lead poisoning in cattle and chickens in the state of Pará, Brazil

The present study describes the occurrence of lead poisoning in cattle and chickens in Pará, Brazil. In a lot composed of 80 calves from a dairy herd, 10 animals became sick and nine died, but one animal recovered after being removed from the paddock. Upon inspection of this paddock, the presence of truck batteries used to store energy captured by solar panels was found. The clinical signs observed in calves included difficult breathing, nasal discharge, excessive salivation, corneal opacity, pushing of the head against objects and recumbency. The chickens had decreased oviposition and produced eggs with thin or malformed shells. The necropsy findings of the cattle, as well as the histopathological changes observed, were of little significance except for one animal that showed mild astrocytosis histopathology in the cerebral cortex. In one of the chickens, renal histopathology showed mild multifocal acute tubular necrosis. The mean lead concentrations in the livers and kidneys of the cattle were 93.91mg/kg and 209.76mg/kg, respectively, and the mean concentration detected in chicken livers was 105.02mg/kg. It was concluded that the source of lead poisoning in these calves and chickens were the truck battery plates, which were within reach of the animals.

Research of Klebsiella pneumoniae in dairy herds

Klebsiella pneumoniae is a common environmental agent of clinical and subclinical mastitis affecting dairy herds, and may be present in the final product decreasing its quality. Mastitis caused by K. pneumoniae is even more severe due to its poor response to antibiotic therapy, rapid evolution to toxic shock and death of the animal. This paper aimed to study the prevalence of this pathogen among dairy herds in ten farms located in different municipalities of São Paulo State based on size and use of milking technology. All mammary glands of all lactating cows were screened using the California Mastitis Test (CMT) and a strip cup. A single aseptic milk sample (20mL) was collected from all CMT-positive quarters and bulk tanks, whereas swab samples were collected from feces, hind limbs of the animals, bedding and milking parlor. Identification of K. pneumoniae was performed using conventional microbiology culture, biochemical assay and Polimerase Chain Reaction. The primers were designed and tested at the Laboratory of Molecular Biology applied to Zoonoses (FMVZ, Unesp-Botucatu) targeting the 16S rRNA gene. This study included 1067 animals. Six cases of intramammary infection by K. pneumoniae were detected in six different cows in two farms. Moreover, K. pneumoniae was isolated in 77 swabs (34 from bedding in 9 farms, 7 from waiting rooms in 5 farms, 6 from milking parlors in 4 farms, 11 from rectums in six farms, and 19 from hindlimbs in 7 farms. Molecular analysis confirmed the agent was K. pneumoniae. At least one strain of the agent was identified in a certain site in all farms, showing the need of maintaining the hygiene in dairy farms.

Histopathological, immunohistochemical, and molecular study of BHV-5 infection in the central nervous system of experimentally infected calves

Bovine meningoencephalitis caused by BHV-5, a double-stranded DNA enveloped virus that belongs to the family Herpesviridae and subfamily Alphaherpesvirinae, is an important differential diagnosis of central nervous diseases. The aim of this study was to describe the histological changes in the central nervous system of calves experimentally infected with BHV-5 and compare these changes with the PCR and IHC results. Formalin-fixed paraffin-embedded central nervous system samples from calves previously inoculated with BHV-5 were microscopically evaluated and tested using IHC and PCR. All the animals presented with nonsuppurative meningoencephalitis. From 18 evaluated areas of each calf, 32.41% and 35.19% were positive by IHC and PCR, respectively. The telencephalon presented more accentuated lesions and positive areas in the PCR than other encephalic areas and was the best sampling area for diagnostic purposes. Positive areas in the IHC and PCR were more injured than IHC and PCR negative areas. The animal with neurological signs showed more PCR- and IHC-positive areas than the other animals.

Recovery of Mollicutes from the reproductive tract of dairy cattle in the state of Pernambuco, Brazil

Abstract: The aim of the present study was to report the occurrence of members of the Mollicutesclass in the reproductive system of dairy cattle in Brazil. Five farms containing dairy cattle were visited in January of 2012. In total, 100 cows of different ages, breeds and stages of lactation were examined in the present study. The cows were part of intensive or semi-intensive management systems and were submitted to mechanical milking or hand milking. The samples were collected after washing the vulvar region with water and soap, and then drying it with paper towels and disinfecting the area with alcohol (70°GL). Vaginal mucous was collected using a sterile alginate cotton swab, which was rubbed on the vagina, as well as the lateral and internal walls. Vulvovaginal mucous samples were cultured in both liquid and solid modified Hayflick´s medium, for mycoplasmas, and UB medium, for ureaplasmas. The PCR assays for Mollicutesand Ureaplasmaspp. were performed according to the standard protocols described in the current literature. During isolation, the frequency of Mycoplasmaspp. was of 13.0% (13/100) and for Ureaplasmaspp. was of 6.0% (6/100). In the PCR assays the frequency of Mollicuteswas of 26.0% (26/100) and for Ureaplasmaspp. was of 13.0% (13/100) in the dairy cattle studied. This is the first report of these agents in reproductive system of bovine of the Pernambuco state. Further studies are necessary to determine the pathogenic potential and species of these field isolates.

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.

The requirements of a feasible plastic packaging material for cultured dairy products

The objective of this Master´s Thesis was to conduct a wide scale preliminary survey regarding the package requirements of a cultured dairy package, and to compare the currently used material polystyrene to other suitable packaging materials. Polystyrene has a long history of use in dairy cups, but in recent years its price has increased significantly compared to other common packaging materials. The overall environmental effects of a package and a package material are today a part of designing a sustainable product life cycle. In addition, in certain contexts there has been discussion of the risks posed by styrene polymer for the environment and for humans. These risks are also discussed in this thesis. Polystyrene (PS) is still the most widely used material in dairy cups. In recent years, polypropylene (PP) cups have appeared in increasing numbers on market shelves. This study focuses on the differences of the suitable polymers and examines the suitability of alternative “suitable” polymers with regards to dairy packaging. Aside from focusing on the cup manufacturer, this thesis also examines its subject matter from the viewpoint of the dairy customer, as well as observing the concrete implications of material changes in the overall value chain. It was known in advance that material permeability would be one of the determining factors and that gas transmission testing would be a significant part of the thesis. Mechanical tests were the second part of the testing process, providing information regarding package strength and protectiveness during the package’s life cycle. Production efficiency, along with uninterrupted stable production, was another important factor that was taken into consideration. These two issues are sometimes neglected in similar contexts due to their self-evident nature. In addition, materials used in production may have a surprising significance to the production and efficiency. Consistent high quality is also partly based on material selection. All of the aforementioned factors have been documented and the results have been analyzed by the development team at Coveris Rigid Finland. Coveris is now calculating the total finance effects and capacities should the material changes be implemented in practice. There are many factors in favor of switching to polypropylene at the moment. The overall production costs, as well as the environmental effects of resin production are the primary influences for said switch from the converters’ perspective.

Synchronization of ovulation in crossbred dairy heifers using gonadotrophin-releasing hormone agonist, prostaglandin F2a and human chorionic gonadotrophin or estradiol benzoate

Girolando (Gir x Holstein) is a very common dairy breed in Brazil because it combines the rusticity of Gir (Bos indicus) with the high milk yield of Holstein (Bos taurus). The ovarian follicular dynamics and hormonal treatments for synchronization of ovulation and timed artificial insemination were studied in Girolando heifers. The injection of a gonadotrophin-releasing hormone (GnRH) agonist was followed 6 or 7 days (d) later by prostaglandin F2a (PGF2a). Twenty-four hours after PGF2a injection either human chorionic gonadotropin (hCG, GPh-d6 and GPh-d7 groups) or estradiol benzoate (EB, GPE-d6 and GPE-d7 groups) was administered to synchronize ovulation and consequently allow timed artificial insemination (AI) 24 and 30 h after hCG and EB injection, respectively. Follicular dynamics in Girolando heifers was characterized by the predominance of three follicular waves (71.4%) with sizes of dominant follicles (10-13 mm) and corpus luteum (approximately 20 mm) similar to those for Bos indicus cattle. In the GnRH-PGF-hCG protocol, hCG administration induced earlier ovulation (67.4 h, P<0.01) compared to the control group (GnRH-PGF) and a better synchronization of ovulation, since most of it occurred within a period of 12 to 17 h. Pregnancy rate after timed AI was 42.8 (3/7, GPh-d6) to 50% (7/14, GPh-d7). In contrast, estradiol benzoate (GnRH-PGF-EB protocol) synchronized ovulation of only 5 of 11 heifers from the GPE-d7 group and of none (0/7) from the GPE-d6 group, which led to low pregnancy rates after timed AI (27.3 and 0%, respectively). However, since a small number of Girolando heifers was used to determine pregnancy rates in the present study, pregnancy rates should be confirmed with a larger number of animals.

Electrophoretic analisys to detect and quantify additional whey in milk and dairy beverages

Polyacrylamide gel electrophoresis, SDS-PAGE system, was adjusted to detect the presence of additional whey in dairy beverages distributed in a Brazilian Government School Meals Program. Aqueous solutions of samples in 8 M urea were submitted to a polyacrylamide gel gradient (10% to 18%). Gel scans from electrophoresis patterns of previously adulterated milk samples showed that caseins peak areas decreased while peak areas of beta -lactoglobulin plus alpha -lactalbumin increased as the percentage of raw milk powder replaced by whey powder increased. The relative densitometer areas of caseins or beta -lactoglobulin plus alpha -lactalbumin plotted against the percentage of whey added to the raw milk showed a linear correlation coefficient square higher than 0.97. The caseins plot was used to determine the percentage of additional whey in 116 dairy beverages, chocolate or coffee flavor. Considering that the lowest relative caseins concentration found in commercial milk powder samples by the present method was 72%, the dairy beverages containing caseins percentages equal to or higher than this value were considered free of additional whey. Based on this criterion, about 49% of the coffee-flavor dairy beverages and 29% of the chocolate-flavor beverages, among all the samples analyzed were adulterated with whey protein to reach the total protein contents specified on their labels. The present method showed a sensitivity of 5% to additional whey.

Avaliação da vida-de-prateleira de bebidas lácteas preparadas com "fat replacers" (Litesse e Dairy-lo)

Face ao aumento de peso e ao aparecimento de doenças cardiovasculares na população mundial devido ao consumo de alimentos altamente calóricos, o desenvolvimento de alimentos com baixo ou reduzido teor de gordura torna-se essencial. Estudou-se o efeito da adição de "fat replacers" (Litesse e Dairy-lo) na vida-de-prateleira de bebidas lácteas. No presente trabalho as variáveis independentes Litesse (X1) e Dairy-lo (X2) foram empregadas nas concentrações 0,50,1,50, 2,50% em peso, com 7 combinações calculadas e 3 repetições do ponto central utilizando a metodologia de superfície resposta. As bebidas lácteas foram avaliadas através de análises químicas (valor de pH, acidez total titulável, teores de sólidos totais e tirosina) e sensoriais (aparência, sabor e consistência) aos 0, 7, 14, 21 e 28 dias de armazenamento a 5ºC. Durante a vida-de-prateleira a utilização de diferentes concentrações de Litesse e Dairy-lo não influenciou o comportamento físico-químico e a aparência das bebidas lácteas estudadas. Entretanto, o sabor foi afetado com o armazenamento. Pode-se estabelecer 28 dias como o tempo ideal para a vida-de-prateleira das bebidas lácteas estudadas. Os resultados indicam que a utilização do Litesse e Dairy-lo em formulações de bebidas lácteas é perfeitamente viável.

Starter cultures used in probiotic dairy product preparation and popular probiotic dairy drinks

Dairy products that contain probiotic bacteria are those that are produced with various fermentation methods, especially lactic acid fermentation, by using starter cultures and those that have various textures and aromas. Fermented dairy products are popular due to their differences in taste and their favourable physiological effects. Today, fermented dairy beverages in general are produced locally by using traditional methods. Recently, due to the increased demand for natural nutrients and probiotic products, fermented dairy beverages have reached a different position and are considered to have an important impact on human health and nutrition. In this article, probiotic bacteria and functional dairy products that are produced by using probiotic bacteria are discussed.