Corrigendum to "Genetic diversity and iron metabolism of Staphylococcus hominis isolates originating from bovine quarter milk, rectal feces, and teat apices" (J. Dairy Sci. 105:9995-10006).

Staphylococcus chromogenes TA showed significantly lower growth under iron-deprived conditions, and adding an iron supplement (lactoferrin or ferritin) resulted in no improvement in growth; in contrast, growth of S. chromogenes IM was significantly recovered with ferritin iron supplementation. OnlyStaphylococcus hominis strains originating from quarter milk were able to significantly utilize ferritin as an iron source to reverse the growth inhibition caused by chelating agent 2,2'-bipyridyl in varying degrees. Both S. chromogenes strains (IM and TA) and all S. hominis strains were unable to significantly use lactoferrin as an iron source for growth recovery.

Comparison of non-aureus staphylococcal and mammaliicoccal species found in both composite milk and bulk-tank milk samples of dairy cows collected in tandem.

Non-aureus staphylococci and the closely related mammaliicoccal species (NASM) are the most common causes of bovine subclinical mastitis on modern dairy farms and are highly prevalent in bulk-tank milk. The purpose of this study was to determine the distribution of NASM in both composite cow milk (CCM) and bulk-tank milk (BTM) samples collected in tandem in commercial Flemish dairy herds and to estimate the origin of the different (subgroups of) NASM species present in BTM by applying strain typing (random amplification of polymorphic DNA or random amplified DNA [RAPD]). A single cross-sectional sampling was performed over 5 herds that volunteered to participate in the study. Composite cow milk samples (n = 356) were collected from all lactating cows (except those with clinical mastitis) during a milking in tandem with 6 BTM samples per herd sequentially collected immediately post that milking (n = 30). In total, 421 and 80 NASM isolates were recovered and identified by MALDI-TOF mass spectrometry from the CCM and BTM samples, respectively and a total of 21 and 12 different NASM species were identified from CCM and BTM samples, respectively. Staphylococcus cohnii was the most prevalent NASM species found in BTM followed by Staphylococcus haemolyticus, Staphylococcus epidermidis, Mammaliicoccus lentus, and Staphylococcus equorum, whereas from CCM samples the most common species were S. hemolyticus, S. cohnii, S. equorum, S. epidermidis, and Staphylococcus chromogenes. The prevalent NASM species in both CCM and BTM samples was distinct for each herd, corroborating other studies observing a herd-specific NASM microbiota. Random amplified DNA analysis was performed on 9 NASM species (S. chromogenes, S. epidermidis, S. haemolyticus, S. equorum, Mammaliicoccus sciuri, Staphylococcus xylosus, S. cohnii, Staphylococcus debuckii, and M. lentus) because these species were isolated from both sample types in a herd. The same RAPD types were found in both sample types for all NASM species selected for strain typing in varying degrees. When assessing the distribution of NASM species, differences within NASM species should be examined meaning a closer look should be taken at the strain level rather than at the species level only.

Genetic diversity and iron metabolism of Staphylococcus hominis isolates originating from bovine quarter milk, rectal feces, and teat apices.

Staphylococcus hominis, a member of the non-aureus staphylococci (NAS) group, is part of the human and animal microbiota. Although it has been isolated from multiple bovine-associated habitats, its relevance as a cause of bovine mastitis is currently not well described. To successfully colonize and proliferate in the bovine mammary gland, a bacterial species must be able to acquire iron from host iron-binding proteins. The aims of this study were (1) to assess the genetic diversity of S. hominis isolated from bovine quarter milk, rectal feces, and teat apices, and (2) to investigate the capacity of bovine S. hominis isolates belonging to these different habitats to utilize ferritin and lactoferrin as iron sources. To expand on an available collection of bovine S. hominis isolates (2 from quarter milk, 8 from rectal feces, and 19 from teat apices) from one commercial dairy herd, a subsequent single cross-sectional quarter milk sampling (n = 360) was performed on all lactating cows (n = 90) of the same herd. In total, 514 NAS isolates were recovered and identified by MALDI-TOF mass spectrometry; the 6 most prevalent NAS species were S. cohnii (33.9%), S. sciuri (16.7%), S. haemolyticus (16.3%), S. xylosus (9.6%), S. equorum (9.4%), and S. hominis (3.5%). A random amplified polymorphic DNA (RAPD) analysis was performed on 46 S. hominis isolates (19 from quarter milk, 8 from rectal feces, and 19 from teat apices). Eighteen distinct RAPD fingerprint groups were distinguished although we were unable to detect the presence of the same RAPD type in all 3 habitats. One S. hominis isolate of a distinct RAPD type unique to a specific habitat (8 from quarter milk, 3 from rectal feces, and 4 from teat apices) along with the quality control strain Staphylococcus aureus ATCC 25923 and 2 well-studied Staphylococcus chromogenes isolates ("IM" and "TA") were included in the phenotypical iron test. All isolates were grown in 4 types of media: iron-rich tryptic soy broth, iron-rich tryptic soy broth deferrated by 2,2'-bipyridyl, and deferrated tryptic soy broth supplemented with human recombinant lactoferrin or equine spleen-derived ferritin. The growth of the different strains was modified by the medium in which they were grown. Staphylococcus chromogenes TA showed significantly lower growth under iron-deprived conditions, and adding an iron supplement (lactoferrin or ferritin) resulted in no improvement in growth; in contrast, growth of S. chromogenes IM was significantly recovered with iron supplementation. Staphylococcus hominis strains from all 3 habitats were able to significantly utilize ferritin but not lactoferrin as an iron source to reverse the growth inhibition, in varying degrees, caused by the chelating agent 2,2'-bipyridyl.

Lymphocyte proliferative responses in dairy cows supplemented with an immunomodulatory feed additive and administered polyvalent vaccination / [Proliferação de linfócitos em vacas leiteiras suplementadas com um aditivo alimentar imunomodulador e vacinadas com vacina polivalente]

O presente estudo avaliou o efeito da suplementação com OmniGen-AF® na proliferação de linfócitos e títulos de anticorpos após vacinação em bovinos leiteiros. Amostras de sangue periférico foram coletadas de 32 vacas leiteiras para quantificação dos títulos de anticorpos anti-Leptospira, e amostras de sangue periférico de 16 vacas leiteiras foram também coletadas para avaliação da proliferação de linfócitos. Observou-se que a suplementação com OmniGen-AF® aumentou a proliferação basal de linfócitos (sem estímulos) 21 dias após a vacinação (P=0,03), apesar de reduzir a proliferação de linfócitos B quando estimulada com Leptospira borgpetersenii serovar Hardjo inativada pelo calor (P=0,03). Ademais, nenhum efeito da suplementação sobre a proliferação de linfócitos no momento imediatamente anterior à vacinação e nos títulos de anticorpos anti-Leptospira foi encontrado. Além disso, a proliferação de linfócitos estimulada com lipopolissacarídeos foi maior em vacas multíparas que em primíparas 21 dias após a vacinação (P=0,03). Desse modo, o presente estudo demonstra que a suplementação com OmniGen-AF® não afetou de forma robusta a proliferação de linfócitos e os títulos de anticorpos anti-Leptospira após vacinação em vacas leiteiras sadias.(AU)

Avaliação da atividade funcional dos neutrófilos sanguíneos em ovelhas primíparas e pluríparas da raça Lacaune / [Functional evaluation of blood neutrophils in primiparous and pluriparous Lacaune ewes]

This study aimed to evaluate the neutrophil oxidative metabolism and phagocytosis of Zymonsan particles of primiparous and pluriparous Lacaune ewes during the first 30 days after lambing. A total of 20 ewes were evaluated, 10 primiparous (GPR) and 10 pluriparous (GPL). Evaluation of basal oxidative metabolism was performed using the nitroblue tetrazolium (NBT) technique, stimulating neutrophil phagocytosis with Zymosan particles. Blood samples were collected at parturition day (M1) and 1, 3, 7, 15 and 30 days after parturition, corresponding to M2, M3, M4, M5 and M6, respectively. In relation to the groups, GPR presented lower oxidative basal metabolism neutrophils in M1 compared to M4, in M3 with M1, M2, M4 and M5. In M4 and M5 differences were found at all times and in M6 with M4 and M5. Higher percentage of neutrophils than phagocytes were found in M4, M5 and M6 than in M1 and M2 in GPL animals. At all times GPR presented a lower percentage of phagocytosis than GPL. Thus it is concluded that the immune response in pluriparous sheep was more efficient than in primiparous sheep.(AU)

Immune response in nonspecific mastitis: What can it tell us?

We analyzed a large number of immune response parameters from quarter milk samples with distinct bacteriological and quarter somatic cell count (qSCC) statuses. Furthermore, we sought to explore and identify displayed immune response patterns in milk samples from mammary glands with nonspecific mastitis. Thus, 92 quarter milk samples from 28 cows were stratified into 4 groups, as follows: (1) 49 culture-negative control quarters with a low qSCC (<1 × 105 cells/mL) from 19 dairy cows (so-called healthy quarters); (2) 15 culture-negative quarters with high qSCC (>2 × 105 cells/mL; so-called quarters with nonspecific mastitis) from 10 dairy cows; (3) 8 culture-positive quarters with low qSCC (noninflammatory quarters with low qSCC) from 5 dairy cows; and (4) 20 culture-positive quarters with high qSCC (so-called truly infected quarters) from 8 dairy cows. Using flow cytometry, we evaluated the percentage of milk neutrophils and their viability, intracellular reactive oxygen species production, phagocytosis, and the expression of CD62L, CD11b, and CD44 for each of the 4 quarter strata. Furthermore, the percentage of monocyte/macrophages, B cells, and T lymphocyte subsets were evaluated by flow cytometry. Milk samples from bacteriologically negative quarters (both with a low and elevated qSCC) had a lower qSCC than those with bacteriologically positive outcomes (both with a low and elevated qSCC). As expected, the healthy quarters showed the lowest percentage of neutrophils and also showed a higher percentage of milk monocytes/macrophages and lower percentage of T lymphocytes than truly infected quarters. The most prominent result of the present study is that quarters with nonspecific mastitis showed the highest percentage of milk CD4+ T lymphocytes. The healthy quarters had a lower percentage of apoptotic neutrophils than noninflammatory and truly infected quarters, although it did not differ from those from the quarters with nonspecific mastitis. Our study supports the role of differential cell counting in the diagnosis of mastitis, as the milk leukocyte populations markedly fluctuate under healthy and inflammatory conditions. Furthermore, an increase in milk CD4+ T cells was associated with nonspecific mastitis, suggesting an increase in this leukocyte subpopulation is correlated with low bacterial shedding. Our study allows us to go further in our understanding of mammary gland immunity, providing further insights on potential protective mammary gland immunity, which we hypothesize can open new avenues for the development of novel targets that can promote bovine udder health.

Genetic parameters and response to selection for growth in tambaqui.

Although the tambaqui (Colossoma macropomum) is the most cultivated native fish species in Brazil, estimated breeding values for growth traits are rarely used for selection of superior individuals in commercial fingerling production. This study aimed to estimate the (co)variance components of growth traits. Body weight, length and width of 2500 tambaqui were determined at tagging and at 6 and 12 months after tagging in a commercial breeding programme in Brazil. Heritability estimates were low for traits measured at tagging (0.10 to 0.19) and moderate to high for traits measured at 6 and 12 months (0.23 to 0.81). Common full-sib effects were high at tagging (>73%), low at 6 months and negligible at 12 months. Positive genetic correlations were found among growth traits at 12 months (0.84 to 0.99) and between growth traits at 6 and 12 months (0.80 to 0.92). These results show that animal selection can be performed at 6 months after tagging. Expected genetic gains for growth traits ranged from 8% to 31%. A simulation of the sex ratio was performed, as individuals did not reach sexual maturity during the experimental period. Because of the sexual dimorphism, more accurate heritability estimates were obtained when considering the female proportion to be 90% in the high-weight group. The findings indicate that it is possible to obtain considerable genetic gains in growth by selecting for growth traits. The development of a tool to determine the sex of animals at early stages can improve the response to selection in tambaqui.

Comparison of antibody repertories against Staphylococcus aureus in healthy and infected dairy cows with a distinct mastitis history and vaccinated with a polyvalent mastitis vaccine.

Staphylococcus aureus is one of the pathogens most frequently isolated from cases of mastitis worldwide. To decrease the effect of S. aureus mastitis in dairy farming, alternative strategies for controlling mastitis are needed that depend on a better knowledge of cow-to-cow variations in S. aureus antibody production. The present study sought to explore the diversity of S. aureus antibodies produced by dairy cows with a distinct mastitis history and vaccinated with a polyvalent mastitis vaccine. We obtained protein extracts from S. aureus isolates derived from persistent subclinical mastitis. Proteins were fractionated using 2-dimensional gel electrophoresis and Western blotting. Then, Western blotting membranes were exposed to sera from 24 dairy cows that had been divided into the following groups: vaccinated dairy cows that were infected with S. aureus, further subdivided according to whether they (a) remained infected by S. aureus or (b) recovered from the intramammary infection; unvaccinated dairy cows infected with S. aureus; and vaccinated healthy dairy cows with no history of S. aureus mastitis. Proteins found to be reactive by Western blot were identified by mass spectrometry (MALDI/TOF-TOF). Our most important finding was that F0F1 ATP synthase subunit α, succinyl-diaminopimelate desuccinylase, and cysteinyl-tRNA synthetase were potential candidate proteins for the prevention of S. aureus mastitis. This study strengthens the notion that variations among animals should not be ignored and shows that the heterogeneity of antibody production against anti-staphylococcal antigens in animals may enable the identification of new immunotherapy targets.

Inhibition of the growth of major mastitis-causing pathogens by non-aureus Staphylococcus isolates using the cross-streaking method / Comunicação: Inibição do crescimento dos principais patógenos causadores de mastite por isolados de estafilococos não aureus pelo método cross-streaking

O objetivo do presente estudo foi avaliar a capacidade de estafilococos não aureus (NAS) isolados de diferentes nichos ecológicos (leite, ambiente e ápice do teto), associados a vacas leiteiras, de inibir os principais agentes etiológicos da mastite bovina (Staphylococcus aureus, Streptococcus agalactiae, Streptococcus uberis e Escherichia coli). Neste estudo, 38 isolados NAS de diferentes nichos ecológicos foram avaliados quanto à capacidade de inibir o crescimento in vitro de importantes patógenos causadores de mastite pelo método cross-streaking. No total, 19 (50%) isolados de NAS (oito isolados de S. chromogenes, 10 de S. fleurettii e um de S. haemolyticus) apresentaram inibição contra os principais patógenos causadores de mastite. No entanto, a inibição dos patógenos causadores da mastite bovina por isolados de NAS foi maior contra bactérias Gram-positivas. Além disso, o presente estudo não sugeriu que os nichos ecológicos influenciam a capacidade do NAS de inibir os principais patógenos causadores da mastite bovina. Com base nesses resultados, concluiu-se que certos isolados de NAS apresentam potencial efeito protetor contra os principais patógenos da mastite, pelo menos in vitro.(AU)

Effect of seasonal conditions and milk management practices on bulk milk quality in Minas Gerais State - Brazil / Efeito da sazonalidade e práticas de manejo na qualidade do leite de tanque no estado de Minas Gerais - Brasil

The influence of seasonality and the training and implementation of good dairy farming practices on raw milk production and quality was evaluated on dairy farms in Minas Gerais State, Brazil. The physico-chemical composition, somatic cell count (SCC) and total bacterial count (TBC) were determined in 3,096 milk samples collected from bulk tanks originated from 43 dairy farms over a three-year period and correlated with key climatic factors. The recommended milking management practices were applied through a training program and correlated with the seasonal data in three stages: I) prior to training (dry period); II) 48 days after the training (transition period); III) 96 days after the training (rainy period). In the first stage, a diagnosis of the situation was performed with raw milk samplings for laboratory analysis, and training for implementation of good milking practices. In stages II and II, the checklist and laboratory analysis were also performed. The rainfall and high temperatures were found to represent the main factors affecting the milk composition and production, and TBC. The composition and physical properties of raw milk, and the TBC and SCC parameters can be controlled or minimized by applying proper milking management practices and constant monitoring.(AU)

Avaliou-se a influência das condições climáticas em regiões tropicais, bem como do treinamento e da implementação de boas práticas de manejo na produção e qualidade do leite em distintos períodos em fazendas leiteiras no estado de Minas Gerais, Brasil. No presente estudo, as características físico-químicas do leite, a contagem bacteriana total (CBT) e a contagem de células somáticas (CCS) foram determinadas em 3.096 amostras de tanques de leite proveniente de 43 fazendas leiteiras, durante um período de três anos, e correlacionadas com os principais fatores climáticos (temperatura diária do ar mínima, média e máxima; pluviosidade e umidade relativa). Foram avaliados os resultados obtidos nas análises das amostras de leite cru coletadas no ano anterior (2009/2010) e também no posterior (2011/2012) àquele em que foi realizado o treinamento para implementação das boas práticas de manejo de ordenha (2010/2011), compreendendo três etapas: antes do treinamento (período seco - tempo 0, etapa I), 48 dias após o treinamento (período de transição - etapa II) e 96 dias após o treinamento (período chuvoso - etapa III). Na etapa I foi realizado um diagnóstico de situação com coleta de amostras para análises laboratoriais, aplicação da lista de verificação padronizada e treinamento para a implementação das boas práticas de manejo de ordenha; na etapa II foram realizadas novas análises laboratoriais e reaplicação da lista de verificação para avaliar a eficiência do treinamento; e na etapa III foi realizada a repetição da etapa II. Com base no histórico dos três anos, observou-se que condições climáticas de alta temperatura e pluviosidade representam importantes fatores que afetam a composição do leite e o volume produzido, assim como parâmetros higiênico-sanitários do leite. Ademais, as boas condições de manejo são ferramentas úteis, eficazes, práticas e essenciais para a maior produção de leite com qualidade, desde que constantemente monitoradas.(AU)

Short communication: Occurrence of methicillin-resistant Staphylococcus aureus and coagulase-negative staphylococci in dairy goat herds in Ohio, United States.

In light of the scarcity of information about the occurrence and epidemiology of methicillin-resistant Staphylococcus aureus (MRSA) and coagulase-negative staphylococci (MRCNS) in small ruminants in general, and particularly dairy goats, we launched this limited-scope study. The findings reported here show the detection of MRSA and MRCNS in goat milk and teat skin samples from dairy goat herds in the state of Ohio. A total of 120 milk samples and 120 teat-swab samples were collected from 5 farms. After conventional isolation and phenotypic characterization of the staphylococci colonies, bacterial isolates were tested by PCR assay targeting the genes nuc to identify Staphylococcus aureus and mecA to detect MRSA and MRCNS. The clonal complexes of MRSA isolates was also determined by multiloccus sequence typing. Fifteen (6.2%) positive S. aureus samples were found in this study: 9 from milk and 6 from teat skin samples. Four (2%) MRSA isolates were detected and, using multiloccus sequence typing genotyping, these were designated to clonal complexes CC133 (n = 2; milk samples) and CC5 (n = 2; teat skin). Three (1.25%) coagulase-negative staphylococci isolates from the teat skin also harbored the mecA gene. Although, the MRSA isolated from milk samples is not a typical human-associated lineage, the CC5 clone isolated from teat skin is a common and widespread clonal complex associated with humans, suggesting that this extramammary niche could be a relevant reservoir of methicillin-resistant staphylococci. Furthermore, the fact that 75% of MRSA were recovered from 1 farm showing poor hygiene practices strengthens the hypothesis that good hygiene practices could be useful to prevent persistence and spread of MRSA at a farm level.

Mastitis in the transition period: identification of potential blood markers / Mastite no período de transição: identificação de potenciais marcadores no sangue

This study identified potential blood markers associated with mastitis in dairy cows under different dry therapies during the transition period, using a logistic regression model. Thirty-four Holstein dairy cows were divided into three groups: untreated controls (13 cows, 42 quarters); animals that received an antimicrobial at drying-off (10 cows, 40 quarters); and animals that were administered an internal teat sealant at drying-off (11 cow, 44 quarters). Blood and quarter milk samples were collected 60 days before the expected day of calving, on the day of calving, and three, seven, 15, 21, and 30 days after calving. Milk samples were submitted for bacteriological analysis and somatic cell count. Blood samples were collected for analyses of the following: the erythrogram and leucogram; plasm fibrinogen concentration; hepatic and renal functions; metabolic profile; serum calcium and phosphorous levels; total serum protein and albumin concentrations. The concentration of total serum proteins was associated with a high somatic cell count. Similarly, the concentrations of total serum proteins and triglycerides were associated to milk bacteriological positive samples during the transition period. Thus, the occurrence of mastitis in dairy cows during the transition period was greater in animals that showed higher concentrations of serum total proteins and triglycerides, in contrast to the use of dry cow therapy.(AU)

O presente estudo identificou potenciais marcadores sanguíneos associados tanto à mastite, durante o período de transição em vacas leiteiras submetidas à antimicrobianoterapia, quanto à utilização do selante interno de teto na secagem por modelo de regressão logística. Trinta e quatro vacas da raça Holandesa foram divididas em três grupos experimentais, a saber: 13 animais (42 quartos mamários) que não receberam tratamento na secagem; 10 animais (40 quartos mamários) tratados por via intramamária com uma bisnaga do antimicrobiano para vaca seca à base de cefalônio anidro após a última ordenha em cada quarto mamário; e 11 animais (44 quartos mamários) que receberam, por via intramamária, uma bisnaga de selante à base de subnitrato de bismuto após a última ordenha em cada quarto mamário. As amostras de leite e sangue foram coletadas 60 dias antes da data prevista do parto, na secagem, e no dia do parto e após três, sete, 15 e 21 dias após o parto. As amostras de leite foram utilizadas para o exame microbiológico e a determinação da contagem de células somáticas. As amostras de sangue foram utilizadas para determinação do eritrograma, leucograma, fibrinogênio plasmático, funções hepáticas e renais, perfil metabólico, proteína total e albuminas séricas e concentração sérica de cálcio e fósforo. A concentração total de proteínas séricas foi associada à alta contagem de células somáticas. Similarmente, a concentração total de proteínas séricas e triglicérides foi associada ao isolamento de patógenos causadores de mastite nas amostras de leite durante o período de transição. Desse modo, conclui-se que vacas leiteiras com concentrações séricas maiores de proteína total e triglicérides têm maior chance de apresentar mastite durante o período de transição; em contraste, o uso da antimicrobianoterapia de vaca seca reduz esse risco.(AU)

Perfil de imunoglobulinas, cura clínica e bacteriológica após diferentes tratamentos para a mastite clínica bovina / Profile of immunoglobulins, clinical and bacteriological cure after different treatment routes of clinical bovine mastitis

O presente estudo objetivou avaliar o perfil das Ig durante os diferentes tratamentos de vacas com mastite clínica. Para isso, 30 vacas com mastite clínica em um quarto mamário foram utilizadas e divididas em três grupos. O primeiro grupo foi composto por 10 animais submetidos ao tratamento combinado com infusão intramamária de 8,5mg de sulfato de cefquinoma após cada ordenha, totalizando três aplicações e administração intramuscular de 2,5mg/kg de enrofloxacina por três dias. O segundo grupo foi composto por 10 animais submetidos ao tratamento intramamário, com infusão intramamária de 8,5mg de sulfato de cefquinoma, após cada ordenha, totalizando três aplicações. O terceiro grupo foi composto por 10 animais submetidos ao tratamento sistêmico, com 2,5mg/kg de enrofloxacina, durante três dias. As amostras de leite foram coletadas de todos os animais antes dos tratamentos (momento 0), no segundo (momento 1), no quinto (momento 2) e no 12º dia (momento 3) após o término dos tratamentos. Estas foram submetidas à contagem de células somáticas, ao California Mastitis Test (CMT), ao exame bacteriológico e à quantificação das IgG1, IgG2, IgA e IgM. O tratamento combinado foi mais eficaz e precoce na taxa de cura clínica, na redução dos escores de CMT e da contagem de células somáticas. Além disso, os resultados do presente estudo demonstraram que as concentrações lácteas das diferentes classes de Ig, apesar de sua importância biológica, não estão relacionadas ao prognóstico da mastite clínica bovina, ou seja, não podem ser consideradas marcadores robustos associados à cura clínica e/ou bacteriológica da infecção intramamária.(AU)

The present study aimed to evaluate the profile of immunoglobins profile, clinical and bacteriological cure after different treatment routes of clinical bovine mastitis. Here, 30 Holstein cows with clinical mastitis in one quarter were uniformly divided into: 10 dairy cows that received 8.5mg of cefquinome sulphate administrated intramammarily during three consecutive milkings and 2.5mg/kg of enrofloxacin administrated parenterally during three consecutive days (Group 1); 10 dairy cows that received 8.5mg of cefquinome sulphate administrated intramammarily during three consecutive milkings (Group 2); and 10 dairy cows that received 2.5mg/kg of enrofloxacin administrated parenterally during three consecutive days (Group 3). Milk samples for somatic cell count, California Mastitis Test (CMT), bacteriological culture and quantification of IgG1, IgG2, IgM and IgA were collected before antimicrobial treatment, and after two, five and 12 days after the antimicrobial treatment. The combined treatment was more effective in the clinical cure rate, reduction of somatic cell count and CMT scores. Furthermore, the results demonstrated that milk concentration of different Igs classes were not related to prognosis of bovine clinical mastitis, and then cannot be considered as robust markers associated with clinical and bacteriological cures.(AU)

Factors associated with microbiological and clinical cure of mastitis in dairy cows / Fatores associados à cura microbiológica e clínica da mastite em vacas leiteiras

709 clinical mastitis cases were analyzed and treated with antimicrobial combination cephalexin-neomycin and the anti-inflammatory prednisolone. A sample of milk was collected to perform a microbiological culture before starting the treatment and 14 days later. Somatic cell count (SCC) was obtained from samples collected on the day of the clinical case (D0), 14 days after (D14) and 28 days after (D28). Of the total, 435 (61.4%) at the D0 exhibited growth of microorganisms. Of the isolated agents, 365 (84%) were Gram-positive, and 66 (16%) were Gram-negative. A clinical cure was achieved in 63% of cases. Bacteriological cure occurred in 75% of cases. Only at D28 after the clinical case a significant SCC reduction was verified. The logistic regression for clinical cure showed significant effects for days in milk and parity (P< 0.05). For bacteriological cure, there were significant effects of Log (SCC) D0; clinical cure and quarter affected (P< 0.05). In the principal component analysis, the Temperature-Humidity Index was associated with reduced clinical cure of clinical mastitis cases.(AU)

Setecentos e nove casos clínicos de mastite foram analisados e tratados com combinação antimicrobiana à base de cefalexina-neomicina e o anti-inflamatório prednisolona. Uma amostra de leite foi coletada para realização de cultura microbiológica antes do início do tratamento e 14 dias depois. A contagem de células somáticas (SCC) foi obtida de amostras coletadas no dia do caso clínico (D0), 14 dias após (D14) e 28 dias após (D28). Do total, 435 (61,4%) no D0 apresentaram crescimento de microrganismos, enquanto em 274 (38,6%) não houve crescimento. Dos agentes isolados, 365 (84%) eram Gram-positivos e 66 (16%) eram Gram-negativos. A cura clínica foi alcançada em 63% dos casos. A cura bacteriológica ocorreu em 75% dos casos. Apenas no D28 verificou-se uma redução significativa na SCC. A regressão logística para a cura clínica mostrou efeitos significativos para dias em lactação e paridade (P<0,05). Para a cura bacteriológica, houve efeitos significativos de Log (SCC) D0; cura clínica e quarto afetado (P<0,05). Na análise do componente principal, o índice de temperatura-umidade foi associado com a redução da cura clínica dos casos clínicos da mastite.(AU)

Factor analysis as a tool to estimate association among individual proteins and other milk components with casein micelle size and cheese yield / Análise fatorial como ferramenta para estimar a associação entre proteínas individuais e outros componentes do leite com o tamanho das micelas de caseína e massa seca de queijo

The present study attempted to identify individual milk proteins and other milk components that are associated with casein micelle size (CMS) and dry matter cheese yield (DMCY) using factor analysis. Here, we used 140 bulk tank milk samples from different farms. Milk composition was determined using a Fourier transform infrared equipament. The individual milk proteins were (αS-casein, ß-casein, κ-casein, ß-lactoglobulin and α-lactoalbumin) measured by their electrophoretic profile. The CMS was estimated by photon correlation spectroscopy, and the DMCY was determined using reduced laboratory-scale cheese production. Factor analysis partitioned the milk components into three groups that, taken together, explain 68.3% of the total variance. The first factor was defined as "CMS", while the second as "DMCY" factor, based on their high loadings. The CMS was positively correlated with protein, casein, non-fat solids and αS-casein and negatively associated with κ-casein and ß-lactoglubulin. DMCY was positively correlated with fat, protein, casein, total solids and negatively correlated with αs-casein. These results indicate that the variation of individual milk proteins may be an important aspect correlated to milk quality and cheese production.(AU)

O objetivo do presente estudo foi avaliar a associação das frações proteicas individuais e de outros componentes do leite com o tamanho das micelas de caseína (TMC) e a produção de matéria seca de queijo (MSQ) utilizando-se análise fatorial. Foram coletadas 140 amostras de leite de tanque provenientes de diferentes fazendas. A determinação da composição do leite foi determinada por espectroscopia no infravermelho com transformação de Fourier. As proteínas individuais (αS-caseína, ß-caseína, κ-caseína, ß-lactoglobulina e α-lactalbumina) foram quantificadas pelo perfil eletroforético. O tamanho médio das micelas de caseína foi analisado pelo princípio de espectroscopia de correlação de fótons e pela produção MSQ a partir do modelo de coagulação do leite em escala reduzida. A análise fatorial delimitou as variáveis em três fatores, que, juntos, responderam por 68,3% da variação total dos dados. No primeiro fator foram observadas as associações mais fortes com o TMC, enquanto no segundo fator as correlações foram mais significativas com a MSQ. O TMC foi associado positivamente com o conteúdo de proteína, caseína, sólidos desengordurados e αS-caseína, e negativamente com κ-caseína e ß-lactoglubulina. MSQ foi associada positivamente com o teor gordura, proteína e caseína total, sólidos totais, e negativamente com o teor de αs-caseína. Esses resultados indicam que a variação quantitativa das proteínas do leite pode ser determinante da qualidade do leite na produção de queijo.(AU)

Immunological implications of bovine leukemia virus infection.

This study examined neutrophil and monocyte functions and the blood lymphocyte profile of naturally BLV-infected cows with or without persistent lymphocytosis (PL). The percentage of neutrophils and monocytes that phagocytosed Staphylococcus aureus was lower in BLV-infected dairy cows, particularly those with PL. The relative percentage of CD44+ monocytes and neutrophils and CD11b expression by neutrophils was also lower in BLV-infected dairy cows with PL. A correlation between the percentage of CD11b+ neutrophils and that produced reactive oxygen species (ROS) was found. Furthermore, the percentage of CD44+ monocytes was positively correlated with the percentage of monocytes that phagocytosed S. aureus and the same phenomenon was observed for neutrophils. In BLV-infected dairy cows, particularly those with PL, inhibition of monocyte and neutrophil apoptosis was observed. Additionally, the percentage of neutrophils producing ROS was lower in BLV-infected cows with PL, in contrast to higher intensity of intracellular production of ROS by monocytes. The result from the lymphocyte immunophenotyping of BLV-infected cows with PL was an increase in B cells, mainly B CD5+ CD11b+, due to the apoptosis inhibition. In conclusion, this study provides novel insight into the implications of BLV infection for cattle, which can include the dysfunction of blood monocytes and neutrophils.

Climate conditions associated with the occurrence of antimicrobial and macrocyclic lactone residues in bulk tank milk / Condições climáticas associadas à ocorrência de resíduos de antimicrobianos e lactonas macrocíclicas em amostras de leite de tanque

The present study aimed to identify the climate condition parameters that are associated with the occurrence of antimicrobial and macrocyclic lactone residues in bulk tank milk using a multivariate principal components analysis (PCA). A total of 132 raw milk samples were collected at dairy farms in Minas Gerais State in Brazil and analyzed for 35 analytes, comprising macrocyclic lactones and antibacterials, using liquid chromatography coupled with mass spectrometry in tandem mode spectrometry. Of the 132 samples, 34 (25.76%) bulk tank milk samples were positive for at least one analyte. PCA showed that antimicrobial residues in bulk tank milk occurred less frequently on days with a higher average temperature, maximum temperature and temperature-humidity index. In contrast, relative humidity was inversely associated with antimicrobial residues in raw milk. The PCA showed that daily milk production was also related to macrocyclic lactone residues, while rainfall showed an inverse association. Thus, some climate conditions, such as average temperature, maximum temperature and temperature-humidity index, can predict the moments with lower risk of occurrence of antimicrobial residues in bulk tank milk, in contrast to relative humidity. Furthermore, the risk of macrocyclic lactone residues in bulk tank milk was higher in months with less rainfall.(AU)

O presente trabalho objetivou identificar fatores climáticos associados à ocorrência de resíduos de antimicrobianos e lactonas macrocíclicas em amostras de leite de tanque por análise multivariada de componentes principais (ACP). Para o presente trabalho, 132 amostras de leite cru foram coletadas em fazendas leiteiras localizadas no estado de Minas Gerais (Brasil) e analisadas por cromatografia líquida de alta eficiência e espectrometria de massas in tandem para detecção de 35 analitos, incluindo antimicrobianos e lactonas macrocíclicas. Das 132 amostras de leite analisadas, detectou-se pelo menos um analito em 34 (25,76%) amostras. A ACP demonstrou que a presença de resíduos de antimicrobianos no leite de tanque ocorreu menos frequentemente nos dias com maior temperatura média, temperatura máxima e índice de temperatura e umidade. Por outro lado, a umidade relativa foi inversamente associada à presença de resíduos antimicrobianos no leite. A ACP demonstrou associação entre a presença de resíduos de lactonas macrocíclicas no leite e a produção diária de leite, e a presença de resíduos de lactonas macrocíclicas ocorreu menos frequentemente nos meses com maiores índices pluviométricos. Dessa forma, conclui-se que alguns índices climáticos, como temperatura média, temperatura máxima e índice de temperatura e umidade, podem predizer períodos com maior risco de ocorrência de resíduos de antimicrobianos, em contraste com a umidade relativa. Além disso, o risco de resíduos de lactonas macrocíclicas no leite de tanque foi maior nos meses com menores índices pluviométricos.(AU)

Achados ultrassonográficos da glândula mamária de cabras naturalmente infectadas com o vírus da artrite encefalite caprina / Ultrasonographic findings of mammary glandof goats naturally infected with caprine arthritis encephalitis virus

A artrite encefalite caprina é uma virose que está distribuída mundialmente nos rebanhos. Na forma mamária, é predominante a mastite crônica, que pode causar assimetria e endurecimento do úbere, mais conhecida como mastite endurativa crônica, levando à redução na produção leiteira. A ultrassonografia é um bom método de avaliação da glândula mamária, pois consiste em uma técnica não invasiva, rápida e de fácil aplicação. Este estudo objetivou avaliar as características ultrassonográficas (escala de cinza, ecogenicidade e homogeneidade) do parênquima mamário de 20 cabras sororreagentes e 20 não sororreagentes para o vírus da artrite encefalite caprina. Observou-se que o diâmetro médio dos linfonodos retromamários de animais sororreagentes (0,72cm) para o vírus da artrite encefalite caprina foi menor (P=0,02) que o dos animais não sororreagentes (0,93cm). Na análise entre as porções do úbere, os valores da escala de cinza do terço distal da porção caudal do parênquima mamário foram maiores nos animais sororreagentes (P<0.0001, lado direito; P=0,02, lado esquerdo). Tais dados sugerem que o aumento na escala de cinza do parênquima mamário das cabras pode estar correlacionado com as alterações crônicas causadas pela artrite encefalite caprina, como a mastite endurativa. Em relação aos aspectos subjetivos de ecogenicidade e homogeneidade, os grupos não apresentaram diferença entre si (P>0,05), exibindo em sua maioria padrões hipoecoicos e heterogêneos. Diante dos resultados, o presente estudo aponta indícios de que a artrite encefalite caprina pode causar alterações na escala de cinza mensuráveis pela ultrassonografia, a qual pode ser utilizada como método auxiliar eficiente e não invasivo no monitoramento e diagnóstico precoce da mastite endurativa.(AU)

The caprine arthritis encephalitis is a virosis with worldwide distribution in livestock and in the mammary form chronic mastitis is prevalent, which can cause assimetric and hardening of the udder, known as indurative chronic mastitis leading to reduction in milk production. The ultrasound is a good method of assessing the mammary gland, because the technique is non-invasive, fast, and easy to apply. This study aimed to evaluate the ultrasonographic features (grayscale, echogenicity, and homogeneity) of mammary parenchyma of 20 seroreacting goats and 20 not seroreacting for caprine arthritis encephalitis virus. It was observed that the average diameter of retromammary lymph nodes of seroreacting animals (0.72cm) to caprine arthritis encephalitis virus was smaller (P= 0.02) than in non seroreacting animals (0.93cm). In analysis between the portions of the udder, the grayscale values of the distal third of the caudal portion of the mammary parenchyma was higher in seroreacting animals (P<0.0001, right side; P= 0.02, left side). Such data suggest that the increase in ultrasonographic grayscale of mammary parenchyma of goats can be correlated to chronic changes caused by caprine arthritis encephalitis, as indurative mastitis. In relation to the subjective aspects of echogenicity and homogeneity, the groups showed no difference between them (P>0.05), displaying mostly heterogeneous and hypoechoic standards. From the results, this study points to evidence that the caprine arthritis encephalitis can cause changes in grayscale measurable by ultrasound, and this can be used as an efficient and non-invasive helper method for monitoring and early detection of indurative mastitis.(AU)

Factors affecting carriage and intensity of infection of Calodium hepaticum within Norway rats (Rattus norvegicus) from an urban slum environment in Salvador, Brazil.

Urban slum environments in the tropics are conducive to the proliferation and the spread of rodent-borne zoonotic pathogens to humans. Calodium hepaticum (Brancroft, 1893) is a zoonotic nematode known to infect a variety of mammalian hosts, including humans. Norway rats (Rattus norvegicus) are considered the most important mammalian host of C. hepaticum and are therefore a potentially useful species to inform estimates of the risk to humans living in urban slum environments. There is a lack of studies systematically evaluating the role of demographic and environmental factors that influence both carriage and intensity of infection of C. hepaticum in rodents from urban slum areas within tropical regions. Carriage and the intensity of infection of C. hepaticum were studied in 402 Norway rats over a 2-year period in an urban slum in Salvador, Brazil. Overall, prevalence in Norway rats was 83% (337/402). Independent risk factors for C. hepaticum carriage in R. norvegicus were age and valley of capture. Of those infected the proportion with gross liver involvement (i.e. >75% of the liver affected, a proxy for a high level intensity of infection), was low (8%, 26/337). Sixty soil samples were collected from ten locations to estimate levels of environmental contamination and provide information on the potential risk to humans of contracting C. hepaticum from the environment. Sixty percent (6/10) of the sites were contaminated with C. hepaticum. High carriage levels of C. hepaticum within Norway rats and sub-standard living conditions within slum areas may increase the risk to humans of exposure to the infective eggs of C. hepaticum. This study supports the need for further studies to assess whether humans are becoming infected within this community and whether C. hepaticum is posing a significant risk to human health.

Climate conditions associated with the occurrence of pyrethroid residues in bulk milk tank / Condições climáticas associadas à ocorrência de resíduos de piretroides em amostras de leite de tanque

The present study attempted to identify climate conditions that are associated with the occurrence of pyrethroid residues using a multivariate principal component analysis (PCA). A total of 132 raw milk samples were collected from dairy farms in Minas Gerais State - Brazil and analyzed for seven analytes using gas chromatography with electron capture detection. Of the 132 milk samples, pyrethroid residues were identified in 14 (10.6%) milk samples, of which 12 (9.1%) and two (1.5%) milk samples had the identification of cypermethrin and deltamethrin, respectively. From those samples, nine (6.8%) milk samples were regarded as non-compliant for cypermethrin with this analyte concentration above the maximum residue limits set by Brazilian legislation. A PCA assessing pyrethroid residues in bulk tank milk demonstrated that the average temperature and the Temperature-Humidity Index were associated with pyrethroids residues in bulk milk tank, although the relative humidity was inversely correlated. Thus, the data analysis indicated that the pyrethrid residues associated with some climate conditions can predict the moments with higher risk of occurrence of pyrethroid residues in bulk tank milk.(AU)

O presente trabalho objetivou identificar fatores climáticos associados à ocorrência de resíduos de piretroides em amostras de leite de tanque por análise multivariada de componentes principais (ACP). Para o presente trabalho, 132 amostras de leite cru foram coletadas em fazendas leiteiras localizadas no estado de Minas Gerais (Brasil) e analisadas por sete analitos por cromatografia gasosa com detector por captura de eletróns para detecção de resíduos de piretroides. Das 132 amostras de leite analisadas, a presença de resíduos de piretroides foi detectada em 14 amostras (10,6%), e em 12 (9,1%) e duas (1,5%) foram detectados cipermetrina e deltametrina, respectivamente. Destas amostras, em nove (6,8%) a concentração de cipermetrina encontrada foi maior que a permitida pela legislação brasileira. A ACP demonstrou que a presença de resíduos de piretroides no leite de tanque ocorreu menos frequentemente nos dias com maior umidade relativa, embora a presença desses resíduos estivesse associada a maior temperatura média e a maior índice de temperatura e umidade. Dessa forma, conclui-se que alguns índices climáticos podem predizer períodos com maior risco de ocorrência de resíduos de piretroides no leite de tanque.(AU)