Diagnosis of clinical bovine mastitis by fine needle aspiration followed by staining and scanning electron microscopy in a Prototheca zopfii outbreak

Biopsy by fine needle aspiration together with microbiological examination and scanning electron microscopy were evaluated in diagnosis of clinical bovine mastitis in a Prototheca zopfii outbreak. Fine needle aspiration was performed in 21 mammary quarters from ten Holstein cows presenting clinical mastitis caused by P. zopfii. The algae were previously identified in the microbiological examination of milk collected from these cows. Material aspirated from these 21 mammary glands was submitted to cytological staining (Gram, Giemsa and/or Shor staining). Fine needle aspiration enabled cytological identification of the algae in these 21 mammary glands, from which P. zopfii was isolated in the milk. Simultaneously, five mammary fragments collected by fine needle aspiration from these 21 mammary glands presenting clinical mastitis were also submitted to microbiological examination. P. zopfii was also isolated from these five fragments. Scanning electron microscopy technique also identified three of these five P. zopfii strains isolated from mammary fragments collected by cytological aspiration. These results suggest that fine needle aspiration may be an alternative method for the diagnosis of clinical mastitis.

Peracute bovine mastitis caused by Klebsiella pneumoniae

Relata-se a ocorrência de graves sintomas de mastite hiperaguda em vaca, causada por Klebsiella pneumoniae, na terceira semana de lactação. Descrevem-se aspectos epidemiológicos, sintomas clínicos, procedimentos de diagnóstico microbiológico, conduta terapêutica e medidas de controle.

Relationship between teat-end condition, udder cleanliness and bovine subclinical mastitis

The aims of the present study were to relate intramammary infection (IMI) occurrence and somatic cell count (SCC) with teat-end condition (TEC) and udder cleanliness (UC). Milk samples from 1931 teats were evaluated according to the presence of IMI and SCC. Scores were applied to teats according to the TEC and to UC. Teats ends with a very rough ring had the largest number of IMI when compared to the other three categories, as well as animals with dirtier udders. The change in a TEC score increased by around 30% the chance of IMI. Also, the chance of the animal developing IMI increased by approximately 47% when the UC score increased. No significant association between both scores and quarter SCC was found. It can be concluded that animals with very rough teat end rings and very dirty udders have a greater predisposition to IMI. (C) 2011 Elsevier Ltd. All rights reserved.

Evaluation of fermented milk containing probiotic on dental enamel and biofilm: In situ study

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Healing process of incisor teeth of diabetic rats replanted after storage in milk

Several local factors that influence the healing process of replanted teeth have been investigated. However, it remains unclear how systemic alterations, such as diabetes mellitus, affect the prognosis of these cases. The purpose of this study was to evaluate the healing process of incisors of non-controlled diabetic rats replanted after storage in bovine long shelf-life (UHT) whole milk. Thirty-two rats were randomly assigned to receive an endovenous injection of either citrate buffer solution (group I - control; n = 16) or streptozotocin dissolved in citrate buffer solution to induce diabetes (group II; n = 16). After confirmation of the diabetic status by analysis of the glycemic levels, the maxillary right incisor of each animal was extracted and immersed in milk for 60 min. The root canals of teeth were then instrumented, and were filled with a calcium hydroxide-based dressing and replanted into their sockets. All animals received systemic antibiotic and were killed by anesthetic overdose 10 and 60 days after replantation. The specimens containing the replanted teeth were removed, fixed, decalcified, and embedded in paraffin. Semi-serial 6-mu m-thick sections were obtained and stained with hematoxylin and eosin for histologic and histometric analyses. The results showed that the connective tissue adjacent to the root surface was less organized in the diabetic animals than in the control animals in both periods; the root dentin was less severely affected by root resorption in the diabetic rats; there were no significant differences between the control and diabetic groups regarding the occurrence of replacement resorption and inflammatory resorption.

Production and characterization of a milk-clotting protease in the crude enzymatic extract from the newly isolated Thermomucor indicae-seudaticae N31 (Milk-clotting protease from the newly isolated Thermomucor indicae-seudaticae N31)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Partial characterization of protease from a thermophilic fungus, Thermoascus aurantiacus, and its hydrolytic activity on bovine casein

Proteases are one of the most important groups of industrial enzymes, with considerable application in the food industry. The aim of this work was to study a novel protease produced by the thermophilic fungus, Thermoascus aurantiacus, through solid-state fermentation (SSF). The enzyme acted optimally at pH 5.5 and 60 degrees C it was stable up to 60 degrees C for 1 h and in the pH range 3.0-9.5. To elucidate the enzyme's proteolytic activity, its hydrolytic profile on bovine casein, an important protein in the food industry, was studied by enzymatic hydrolysis on skim milk, analyzed by gel electrophoresis (UREA-PAGE), which clearly showed that the protease does not have the same specificity as bovine chymosin. (c) 2006 Elsevier Ltd. All rights reserved.

AN IMPROVED ELECTROPHORETIC METHOD FOR THE DETERMINATION OF SERUM MILK PROTEIN VARIANTS IN GYR-HOLSTEIN COWS

Milk serum proteins such as alpha-lactalbumin (ALA) and beta-lactoglobulin (BLG) present biochemical polymorphism which is under the control of codominant autosomal alleles. In the present report, we propose modifications of traditional electrophoretic techniques such as increasing the running gel concentration from 5 to 10% and the addition of 5 M urea to the stacking gel, which permitted the detection of two variants (A and B) at the ALA and BLG loci. About 8 mul of milk serum (6 mg/ml protein) and 10 pl of total fresh milk were applied. Bovine serum albumin (BSA) and immunolactoglobulins (ILG) could also be discriminated. Total fresh milk was as useful as the purified serum milk proteins for the discrimination of ALA and BLG serum milk protein polymorphism by alkaline vertical slab polyacrylamide gel electrophoresis. However, BSA and ILG ran with caseins, which prevented their characterization in this system.

Genetic Parameters For the Somatic Cells Count In the Milk of Buffaloes Using Ordinary Test Day Models

The buffaloes dairy milk production (BDMP) has increased in the last 20 years, mainly for the manufacturing of mozzarella cheese, which is recognized by its high nutritional quality. However, this quality can be affected by several factors i. e. high somatic cells count (SCC) provokes changes in the milk's constituents. As in bovine dairy milk, the SCC is used as diagnostic tool for milk quality; because it enables the diagnosis of sub-clinic mastitis and also allows the selection of individuals genetically resistant to that disease. Based on it, we collected information about SCC and BDMP along the lactation in Murrah breed buffaloes, during the period between 1997 and 2005. Curves were designed to estimate genetic parameters. These parameters were estimated by ordinary test-day models. There were observed variations in the estimated heritability for both characteristics the lowest score for somatic cells count (SSCC) was seen at first month (0.01) and the highest at sixth months (0.29 the genetic correlation between these traits varied from -1 at the 1 and 9(th) months to 0.31 and 0.30 in the2 and 4(th) month of lactation. Phenotypic correlations were all negative (-0.07 in the second month and up to -0.35 in the eighth month of lactation). These results showed that environmental factors are more important than genetics in explain SCC, for this reason, selection for genetic resistance to mastitis in buffalos based in SCC should not be done. In the other hand, negative phenotypic correlations demonstrated that as the SCC increased, the milk production decreased.

Productive and reproductive performance in cattle infected with bovine leukosis virus

Economic losses caused by enzootic bovine leukosis (EBL) have been of interest since World War II, when the neoplastic form of EEL increased dramatically in Europe. Olson (1974) and House et al. (1975) ed that animals with lymphosarcoma caused by the bovine leukosis virus (BLV) had reduced milk yields. a less efficient reproductive performance and high veterinary costs and mortality rates, while many carcasses were rejected at slaughter. However, the actual impact of BLV infection in cattle without lymphosarcoma is not; clear. The purpose of the study reported here was to compare some productive and reproductive responses of cattle that were antibody-positive (BLV+) or negative (BLV-) for BLV.Holstein dairy cows in commercial dairy farms were used in this study. Blood samples were collected and subjected to BLV serological examination by the agar gel immunodiffusion test of Miller & van der Maaten (1976). Animals were then grouped as BLV+ or BLV- according to their serological response to the BLV antigen. Productive and reproductive histories were obtained from individual animal records and the following factors were considered: milk production, calving interval and birth rate. For milk production, we had the daily milk yields of 547 animals, and for calving interval the time between two successive parturitions for 444 cows. These values were examined by ANOVA and when this was significant a Student's t test was carried out for each age group. Birth rates, the percentage of animals that calved in 1 gear, were available for 557 animals and were examined with the Z-two proportion test. For all analyses, P < 0.05 was considered significant.

O-serogroups, eae gene and EAF plasmid in Escherichia coli isolates from cases of bovine mastitis in Brazil

Mastitis has been recognized for some time as the most costly disease in dairy herds. From March 1997 to August 1998. 2144 samples of bovine mastitic milk were collected, from which 182 Escherichia coli isolates were made, and from which 14 1 isolates had the somatic anti-en (serogroup) determined. Twelve different serogroups were isolated from mastitic milk, and among them were 026, 055, 0111 and 0 119, all of them classic enteropathogenic E. (oh (EPEC) serogroups. These represented 40.0% of the isolates. The 20 of 57 isolates tested had plasmids and in dot blot hybridization, nine isolates were positive for an EaeA probe and an EPEC adherence factor (EAF) probe while two isolates were negative for EaeA probe but positive for the EAF probe, the nine isolates were characterized as attaching and effacing (A/E) E, coli (AEEC) isolates. (C) 2002 Elsevier B.V. B.V. All rights reserved.

Minimum inhibitory concentrations of Staphylococcus aureus recovered from clinical and subclinical cases of bovine mastitis

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Grazing time and milk production of crossbred cows in a rotational area of Elephant grass and Tanzania grass

This study was carried out in January of two consecutive years. The objective was to evaluate the grazing time, the grazing rate and the milk production of crossbred cows, managed in two rotational grazing areas, one with elephant grass (Pennisetum purpureum Schum. cv. Guaçu) and another one with Tanzania grass (Panicum maximum Jacq. cv. Tanzania), both with natural shade in the rest area. The experiment was divided in two phases, the first with 12 cows that remained day and night in the paddock and were milked twice a day. In the second phase 15 cows were observed and remained 10.43 hours in the paddock during the day, and were kept in a corral during the night. Each cow was observed every 15 minutes. Grass specie had no effect on milk production in the two phases of the experiment. The grazing time was 564 and 474 minutes and the grazing rate was 28.7 and 24.4 minutes/hour for the Elephant grass and Tanzania grass, respectively, in first phase. In the second phase,the grazing time was 461 and 426 minutes and the grazing rate was 42.7 and 39.4 minutes/hour for the Elephant grass and Tanzania grass, respectively. Correlations were observed (P<0.01) between grazing rate and the minimum temperature (-0.68), the maximum temperature (-0.76), the relative humidity (0.44) and the THI (-0.76).

Evaluation of some properties of fermented milk beverages that affect the demineralization of dental enamel

The aim of this in vitro study was to evaluate the erosive capacity of fermented milk beverages, as well as some of their properties that affect the demineralization of dental enamel (pH, buffering capacity, fluoride, calcium and phosphorus contents). Three different batches of 6 commercial brands of fermented milk beverages were analyzed. pH evaluation was accomplished using a potentiometer. The buffering capacity was measured by adding 1 mol L -1 NaOH. Fluoride concentration was assessed by an ion specific electrode after hexamethyldisiloxane-facilitated diffusion, and calcium and phosphorus concentrations were assessed by a colorimetric test using a spectrophotometer. Sixty specimens of bovine enamel were randomly assigned to 6 groups (n = 10). They were exposed to 4 cycles of demineralization in the fermented milk and remineralization in artificial saliva. Enamel mineral loss was determined by surface microhardness (%SMHC) and profilometric tests. The samples' pH ranged from 3.51 to 3.87; the buffering capacity ranged from 470.8 to 804.2 μl of 1 mol L -1 NaOH; the fluoride concentration ranged from 0.027 to 0.958 μgF/g; the calcium concentration ranged from 0.4788 to 0.8175 mgCa/g; and the phosphorus concentration ranged from 0.2662 to 0.5043 mgP/g. The %SMHC ranged from-41.0 to -29.4. The enamel wear ranged from 0.15 μm to 0.18 μm. In this in vitro study, the fermented milk beverages did not promote erosion of the dental enamel, but rather only a superficial mineral loss.

Bovine abortion associated with Neospora caninum: Diagnosis and epidemiological aspects of a dairy cattle herd in the northeast region of São Paulo state, Brazil

The objective of this study was to report the presence of Neospora caninum-associated abortion in bovines at a farm in the northeast region of São Paulo State. In January 2010, it was sent to the Department of Pathology, UNESP-Jaboticabal, a bovine fetus with an estimated age of seven months, which was natural of a dairy farm with 300 animals and an average daily production of 3,000 liters of milk, nearly 20 liters per cow. The animals were vaccinated against rabies, foot and mouth disease, carbuncle, brucellosis, leptospirosis, bovine herpes virus type I and bovine viral diarrhea virus. The herd consisted of purebred Holstein animals, Jersey, and mostly by crossbred animals 7-8 (gir x holstein). During necropsy, samples of the serosanguineous liquid present at the thoracic cavity and the heart of the fetus were collected for the detection of anti-Neospora caninum antibodies through Indirect Immunofluorescence Assay (IFA). Fragments of brain, cerebellum, tongue, liver, heart and kidneys were collected for the execution of histopathology (HP), immunohistochemistry (IHC) and Polymerase Chain Reaction. In order that IFA could be performed, the owner was requested blood samples without anticoagulants of the mother and other cows in the farm, with or without a history of abortion. At necropsy, it was verified a severe autolysis of the fetus. The serology of the fetus was 1:25, while the serology of the mother was 1:3,200. At HP, it was observed discrete multifocal non-suppurative encephalomyelitis characterized by gliosis and mononuclear inflammatory infiltration associated with cellular debris. DNA amplification of N. caninum was positive in fragments of brain, tongue, cerebellum, heart and kidneys. At IHC, it has been observed immunoreactivity to a cyst located in the tongue. The owner reported that his herd showed endemic episodes of abortion, while 27.69% (18/65) of the 65 animals sampled were seropositive. Although it has not been a significant difference (p>0.05), a higher seropositivity was observed in animals with a history of abortion (10/26) 38.46%, in comparison with animals without previous abortion (8/39) 20.51%. These findings show that the abortion under study was provoked by the protozoan N. caninum, while this is the first report concerning cattle in the northeast region of São Paulo State.