Investigation of campylobacteriosis and genital trichomoniasis in bulls from rural properties in the Pantanal of Mato Grosso state, Brazil / Investigação de Campilobacteriose e Tricomonose genital em touros de propriedades rurais do Pantanal Mato-Grossense, Brasil

Bovine genital campylobacteriosis (BGC) and bovine trichomoniasis (BT) are diseases of cattle that are primarily transmitted through sexual contact. Although bulls may be asymptomatic, these infectious diseases contribute to reproductive failure, embryonic death, and abortion in cows. Infection in cattle causes significant economic losses. BGC is caused by two bacterial subspecies, Campylobacter fetus subsp. venerealis and Campylobacter fetus subsp. fetus, whereas the protozoan Tritrichomonas foetus causes BT. Mato Grosso state has the largest bovine herd in Brazil, particularly in the Pantanal region. This area encompasses vast expanses of land characterized by annual floods and a predominant reliance on natural breeding for animal reproduction. These conditions create a favorable environment for the occurrence of BGC and BT within the herd. Given the lack of up-to-date data regarding the prevalence of these diseases, this study aimed to examine the presence of Campylobacter spp. and Tritrichomonas foetus in samples from 100 bulls in the municipalities of Poconé, Santo Antônio de Leverger, and Nossa Senhora do Livramento located in the Pantanal region of Mato Grosso state. Preputial smegma samples were retrieved using preputial swabs and stored in a saline solution at -80ºC for subsequent analysis. Polymerase chain reaction was used to identify the presence of Campylobacter fetus subsp. venerealis, Campylobacter fetus subsp. fetus, and Tritrichomonas foetus. Despite a questionnaire revealing epidemiological conditions conducive to the persistence and spread of these pathogens, they were not detected in the bulls evaluated on rural properties in the Pantanal of Mato Grosso region.(AU)

Campilobacteriose genital bovina (CGB) e Tricomonose bovina (TB) são doenças infectocontagiosas de transmissão venérea, assintomáticas nos touros, sendo consideradas como importantes enfermidades causadoras de falha reprodutiva, morte embrionária ou abortamento, ocasionando perdas econômicas significativas em rebanhos bovinos infectados. CGB é causada pela bactéria Campylobacter fetus subsp. venerealis e Campylobacter fetus subsp. fetus, e TB pelo protozoário Tritrichomonas foetus. O estado de Mato Grosso é detentor do maior rebanho bovino do Brasil, envolve a região do Pantanal Mato-Grossense que possui grandes extensões de terra, com ciclo anual de enchentes e a reprodução dos animais realizada predominantemente por monta natural, condições estas, favoráveis a presença de CGB e TB no rebanho. Considerando a carência de informações recentes sobre a ocorrência dessas enfermidades no estado de Mato Grosso, o objetivo deste estudo foi investigar a presença de Campylobacter spp. e Tritrichomonas foetus em 100 touros provenientes dos municípios de Poconé, Santo Antônio de Leverger e Nossa Senhora do Livramento, localizados na região pantaneira do estado de Mato Grosso. Amostras de esmegma prepucial foram coletadas por meio de escarificação via swab prepucial e armazenadas em solução salina a -80ºC. Para a detecção de Campylobacter fetus subsp. venerealis, Campylobacter fetus subsp. fetus, e Tritrichomonas foetus, foi realizada a reação em cadeia pela polimerase (PCR). Apesar do questionário aplicado nas propriedades revelar condições epidemiológicas que favorecem a manutenção e disseminação desses patógenos, este estudo não identificou a presença dos referidos agentes em touros avaliados nas propriedades rurais do pantanal Mato-Grossense.(AU)

Pseudomonas fluorescens and Pseudomonas putida from refrigerated raw milk: genetic diversity and lipoproteolytic activity.

In this research communication the genetic diversity of Pseudomonas fluorescens (n = 67) and Pseudomonas putida (n = 44) isolated from refrigerated raw milk from bulk tank trucks were verified. The relationship between the genetic profile of the isolates and their lipoproteolytic potential was evaluated using skim milk agar and tributyrin agar (21°C/72 h). The lipoproteolytic potential (low or high), evaluated by the diameter of the halos (cm), was correlated with the number of milk producing properties that contributed to each sample (one sample = one bulk tank truck; 8-80 producers/sample) and the distance between the dairy properties and the processing plant (21-370 km). P. fluorescens was confirmed in all samples, while P. putida in 60% samples. For both species, two clusters (I and II) were observed, and the first one showed lower genotypic diversity and the presence of isolates with 100% similarity. P. fluorescens isolates presenting at least 70% similarity were 83.9% in Cluster I (n = 31) and 44.4% in Cluster II. In both clusters (I and II) observed in the P. fluorescens dendrogram, the occurrence of high proteolytic and lipolytic potential were equivalent. The higher the number of farms per milk sample, the greater the lipoproteolytic intensity of P. fluorescens isolates. In relation to P. putida isolates, 74% presented at least 50% similarity in Cluster I (n = 27) and only 35% in Cluster II (n = 17). The occurrence of high proteolysis linked to P. putida was proportional between both Clusters, but the occurrence of high lipolysis was greater in Cluster II. No significant association was detected between P. putida isolates and the variables studied. The results indicate the circulation of P. putida and P. fluorescens with 100% similarity in different milk producing regions. The level of genetic diversity was related only to the lipolytic capacity of P. putida.

Presence of aprX gene in Pseudomonas spp. from refrigerated raw milk and their proteolytic ability / Presença do gene aprX em Pseudomonas spp. isolados de leite cru refrigerado e sua capacidade proteolítica

This study aimed to determine the frequency of Pseudomonas fluorescens, P. putida, and P. aeruginosa in refrigerated raw milk; their proteolytic potential; and your association with the aprX gene. Of the 173 isolates confirmed as belonging to Pseudomonas spp., 37% were P. fluorescens, 25.4% P. putida, and none belongs to P. aeruginosa. The aprX gene was distributed proportionally between P. putida (68%) and P. fluorescens (75%), but it was not associated with low or high proteolytic potential in both species. P. putida (16) and P. fluorescens (14) isolates with no aprX gene identified also had proteolytic potential. Considering the synthesis of proteases other than AprX by the isolates under study, we concluded that P. fluorescens and P. putida represented 62.4% of the Pseudomonas genus, with high probability of having the aprX gene and proteolytic potential. However, there was no association between the deteriorating potential with the presence of the aprX gene.(AU)

O objetivo do estudo foi verificar a frequência, em leite cru refrigerado de Pseudomonas fluorescens, P. putida e P. aeruginosa, o potencial proteolítico dos isolados e sua associação com a presença do gene aprX. Dos 173 isolados confirmados como pertencente ao gênero Pseudomonas spp, 37% foram P. fluorescens, 25,4% P. putida e nenhum pertencia à espécie P. aeruginosa. A prevalência do gene aprX se distribuiu proporcionalmente entre P.putida (68%) e P.fluorescens (75%) porém não foi associada ao baixo ou alto potencial proteolítico nas duas espécies avaliadas. P. putida (16) e P. fluorescens (14) que não tiveram o gene aprX identificado também expressaram potencial proteolítico, considerando-se a síntese de outras proteases além da AprX pelos isolados estudados. Concluiu-se que Pseudomonas fluorescens e P. putida representaram 62,4 % do gênero Pseudomonas ssp., com alta probabilidade da presença do gene aprX e potencial proteolítico, porém sem associação da expressão do potencial deteriorante com a presença do gene aprX.(AU)

Occurrence and seasonality of aflatoxin M1 in milk in two different climate zones.

BACKGROUND: Aflatoxin M1 (AFM1) is a mycotoxin found in milk as a result of the ingestion of aflatoxin B1 (AFB1) by dairy cattle. Because of its carcinogenic properties, the control of its occurrence in dairy products is of great importance. We evaluated the occurrence and seasonality of AFM1 in milk from farms with subtropical and temperate climates, where significant milk production occurs. Forty samples of raw milk were collected from bulk tansk milk during the summer (five samples from a subtropical climate and 14 from a temperate climate) and winter (six samples from a subtropical climate and 15 from a temperate climate) months of 2017 and conducted an analysis using an enzyme-linked immunosorbent assay (ELISA) (detection limit 5 ng L-1 ). Data from eight random samples were compared with respect to liquid chromatography with mass spectrometry detection and ELISA. A significant correlation was observed. RESULTS: The presence of AFM1 was detected in 87.50% (n = 35) of the samples analyzed (a mean of 16.66 ng L-1 and a median of 12.42 ng L-1 in positive samples); however, its concentration in all samples was below the maximum limit allowed by European (50 ng L-1 ) and Brazilian (500 ng L-1 ) legislations. There were no significant differences in the levels of AFM1 between the properties located in the two climate zones, in both summer and winter. The estimated daily intake of AFM1 based on the milk analyzed was 0.0107 ng kg-1 day-1 for adolescents, 0.0072 ng kg-1 day-1 for adults and 0.0098 ng kg-1 day-1 for the elderly. CONCLUSION: The present work demonstrated a low exposure to AFM1. © 2018 Society of Chemical Industry.

Changes occurring in stabilized ultra-high-temperature-treated whole milk during storage.

This Research Communication describes the relationship between casein, free fatty acids (FFAs) and the storage period of ultra-high temperature-treated (UHT) whole milk observed for a period of 120 d of labelled shelf-life. Moreover, we aim to estimate the daily rate of casein degradation in UHT whole milk, and the total length of time estimated for its full degradation. With this aim, ten sets of samples were evaluated from batches of UHT milk manufactured by a dairy processing plant in Parana State, Brazil on 10 different days. Each set was comprised of one liter of raw milk and 12 units of 1 litre cartons of UHT milk, and represented one batch of production. Total mesophilic (TMC), psychrotrophic (TPC), and somatic cell counts (SCC) of raw milk were assessed. UHT milk was assessed for fat (%), sialic acid (mg/l), casein (%), and FFA contents. TMC ranged from 3·5 × 106 to 3·1 × 107 CFU/ml; TPC, from 106 UFC/ml and higher; and SCC, from 18 × 104 SC/ml to 4·83 × 105 CS/ml. Casein (r = -0·991; R2 = 0·9822) and FFA (r = 0·962; R2 = 0·9245) contents, and storage time of UHT milk were correlated (P < 0·05). The rate of casein hydrolysis was estimated as 0·021 g/100 g UHT whole milk/day. A complete breakdown of casein was estimated to occur by the 560th day post-manufacture. Although age gelation was not observed in our study, the report herein corroborates the understanding that the microbiological quality and SCC of raw milk are important components involving the integrity of casein and lipids of UHT milk during shelf-life.

Proteolytic behavior of isolated Pseudomonas spp. from refrigerated raw milk in different concentrations and storage temperatures / Comportamento proteolítico de Pseudomonas spp. isoladas de leite cru refrigerado em diferentes concentrações e temperaturas de estocagem

The objective of the work was to evaluate the multiplication capacity and proteolytic activity of different Pseudomonas spp. cell counts inoculated in milk and storaged under different temperature. Strains isolated from refrigerated raw milk (RRM) were confirmed at genus level by Polymerase Chain Reaction (PCR). The Pseudomonas spp. was cultured in cephalothin-sodium fusidate-cetrimide (CFC) agar-base (30?C for 48 h) until it reached 2 log and 6 log CFU mL-1. Three of eight strains confirmed as Pseudomonas spp were inoculated in sterile reconstituted whole milk powder and incubated at 2C, 4C, and 8C for 96 h. Primary proteolysis indices was determined by the Kjeldahl method. When taking into account the effect of storage time in Pseudomonas spp. population, it was found that the initial population (2 log CFU mL-1) showed significant difference in growth rates only from 0 h to 24 h, keeping at the same levels along 96 h. When a higher initial population was incubated (6 log CFU mL-1), it was not observed a significant difference for times tested. Related to the effect of storage time in proteolysis index, it was not observed a significant difference in samples inoculated with 2 and 6 log CFU mL-1 Pseudomonas spp. When we analyzed the influence of storage temperature on the bacterial multiplication, there was a significant difference in the Pseudomonas spp. population...

O objetivo deste trabalho foi avaliar a capacidade de multiplicação e a atividade proteolítica de diferentes populações de Pseudomonas spp. inoculadas em leite e estocadas em diferentes temperaturas. Cepas isoladas de leite cru refrigerado (LCR) tiveram o gênero confirmado pela Reação em Cadeia da Polimerase (PCR). Pseudomonas spp. foram isoladas em ágar pseudomonas adicionado de cephalothin-sodium fusidate-cetrimide (30?C por 48 h) até atingir as populações de 2 log e 6 log UFC mL-1. Três de oito cepas confirmadas como Pseudomonas spp. foram inoculadas em leite em pó integral reconstituído esterilizado e incubadas a 2C, 4C e 8C por 96 h. O índice de proteólise primária foi determinado pelo método de Kjeldahl. Considerando o efeito do tempo de estocagem na multiplicação de Pseudomonas spp., observou-se que nas baixas populações iniciais (2 log CFU mL-1) houve diferença significativa na multiplicação bacteriana apenas de 0 h para 24 h, mantendo-se estável ao longo das 96 horas de estocagem. Quando uma população inicial mais alta (6 log UFC mL-1) foi estocada, não observou-se diferença significativa entre o tempo zero e os demais testados. Em relação ao efeito do tempo de estocagem no índice de proteólise, não foi encontrada diferença significativa neste índice, nas amostras com inóculo inicial de 2 e 6 log UFC mL-1 de Pseudomonas spp. Quando foi analisado o efeito da...

Proteolytic behavior of isolated Pseudomonas spp. from refrigerated raw milk in different concentrations and storage temperatures / Comportamento proteolítico de Pseudomonas spp. isoladas de leite cru refrigerado em diferentes concentrações e temperaturas de estocagem

The objective of the work was to evaluate the multiplication capacity and proteolytic activity of different Pseudomonas spp. cell counts inoculated in milk and storaged under different temperature. Strains isolated from refrigerated raw milk (RRM) were confirmed at genus level by Polymerase Chain Reaction (PCR). The Pseudomonas spp. was cultured in cephalothin-sodium fusidate-cetrimide (CFC) agar-base (30?C for 48 h) until it reached 2 log and 6 log CFU mL-1. Three of eight strains confirmed as Pseudomonas spp were inoculated in sterile reconstituted whole milk powder and incubated at 2C, 4C, and 8C for 96 h. Primary proteolysis indices was determined by the Kjeldahl method. When taking into account the effect of storage time in Pseudomonas spp. population, it was found that the initial population (2 log CFU mL-1) showed significant difference in growth rates only from 0 h to 24 h, keeping at the same levels along 96 h. When a higher initial population was incubated (6 log CFU mL-1), it was not observed a significant difference for times tested. Related to the effect of storage time in proteolysis index, it was not observed a significant difference in samples inoculated with 2 and 6 log CFU mL-1 Pseudomonas spp. When we analyzed the influence of storage temperature on the bacterial multiplication, there was a significant difference in the Pseudomonas spp. population...(AU)

O objetivo deste trabalho foi avaliar a capacidade de multiplicação e a atividade proteolítica de diferentes populações de Pseudomonas spp. inoculadas em leite e estocadas em diferentes temperaturas. Cepas isoladas de leite cru refrigerado (LCR) tiveram o gênero confirmado pela Reação em Cadeia da Polimerase (PCR). Pseudomonas spp. foram isoladas em ágar pseudomonas adicionado de cephalothin-sodium fusidate-cetrimide (30?C por 48 h) até atingir as populações de 2 log e 6 log UFC mL-1. Três de oito cepas confirmadas como Pseudomonas spp. foram inoculadas em leite em pó integral reconstituído esterilizado e incubadas a 2C, 4C e 8C por 96 h. O índice de proteólise primária foi determinado pelo método de Kjeldahl. Considerando o efeito do tempo de estocagem na multiplicação de Pseudomonas spp., observou-se que nas baixas populações iniciais (2 log CFU mL-1) houve diferença significativa na multiplicação bacteriana apenas de 0 h para 24 h, mantendo-se estável ao longo das 96 horas de estocagem. Quando uma população inicial mais alta (6 log UFC mL-1) foi estocada, não observou-se diferença significativa entre o tempo zero e os demais testados. Em relação ao efeito do tempo de estocagem no índice de proteólise, não foi encontrada diferença significativa neste índice, nas amostras com inóculo inicial de 2 e 6 log UFC mL-1 de Pseudomonas spp. Quando foi analisado o efeito da...(AU)

Milk adulteration with acidified rennet whey: a limitation for caseinomacropeptide detection by high-performance liquid chromatography.

BACKGROUND: High-performance liquid chromatography (HPLC) is widely employed to determine the caseinomacropeptide (CMP) index and to detect milk tampering with rennet whey. Prior to HPLC analysis, CMP is subject to a trichloracetic acid isolation, causing further soluble proteins in the sample to precipitate. On this basis, we aimed to determine whether rennet whey acidification could adversely affect the HPLC sensitivity with respect to detecting this peptide. RESULTS: As hypothesized, the CMP index from milk with added acidified rennet whey was, on average, half that quantified from milk with added rennet whey. Moreover, the quantum satis of acidified whey added to milk sufficient to demonstrate a HPLC CMP > 30 mg L-1 was 94% greater than that required for this threshold to be reached with rennet whey. CONCLUSION: Milk tampering with acidified rennet whey may limit the analytical sensitivity of the reversed-phase HPLC employed for the screening of CMP and, ultimately, disguise the fraudulent addition of whey to milk. © 2017 Society of Chemical Industry.

Pseudomonas spp. and P. fluorescens: population in refrigerated raw milk / Pseudomonas spp. e P. fluorescens: população em leite cru refrigerado

Raw milk samples were collected from cooling tanks (after they cooled for 48 h) in five dairy farms and the corresponding bulk tank (bulk milk transportation, BMT) when they arrived to the industry. Routine physical chemical analyzes and quantification of psychrotrophic (Pseudomonas spp. and P. fluorescens) and aerobic mesophilic (AM) populations were performed. Only relative density and titratable acidity values for samples of milk from three farms were in agreement to the quality parameters required by law. In the BMT, only the protein content has not reached the minimum value established by law, and counting was performed for AM (>105 colony forming units (CFU) mL-1) and psychrotrophic bacteria (2.8x106CFU mL-1). Pseudomonas spp. counting corresponded to 17.9% of the psychrotrophic population, and P. fluorescens was 3.4% of Pseudomonas spp . count. In milk samples from dairy farms, counts were variable for AM (3.4x105 to 3.7 x107CFU mL-1), psychrotrophic (4.0x104 to 3.1x106CFU mL-1), Pseudomonas spp. (2.3x104 to 1.8x105CFU mL-1), and P. fluorescens (62 to 8.4x103CFU mL-1). For the populations studied, no statistical difference (P>0.05) was observed between counts reported in milk samples collected in dairy farms (cooling tanks) and BMT. Therefore, the genera Pseudomonas spp. and P. fluorescens were not the most frequent psychrotrophic bacteria in this studied milk transportation line.

Amostras de leite foram coletadas de tanques de refrigeração (após 48h de refrigeração) em cinco fazendas de produção e do respectivo caminhão tanque (leite de conjunto, LC) ao chegar à indústria beneficiadora. Análises físico químicas de rotina e quantificação das populações de bactérias psicrotróficas (Pseudomonas spp. e P. fluorescens) e aeróbios mesófilos (AM) foram realizadas. Apenas os valores de densidade relativa e acidez titulável de amostras de leite de três fazendas estavam de acordo com a legislação. No LC, apenas o teor de proteína não atingiu o valor mínimo estabelecido por lei, e a contagem foi realizada para AM (>105 unidades formadoras de colônias (UFC) mL-1) e psicrotróficos (2,8x106UFC mL-1). A contagem de Pseudomonas spp. correspondeu a 17,9% da população de psicrotróficos, e 3,4% da contagem de Pseudomonas spp. eram da espécie P. fluorescens . Nas amostras de leite das fazendas de produtoras, as contagens foram variáveis para AM (3,4x105a 3,7x107UFC mL-1), psicrotróficos (4,0x104 a 3,1x106UFC mL-1), Pseudomonas spp. (2,3x104 a 1,8x105UFC mL-1) e P. fluorescens (62 a 8,4x103UFC mL-1). Para as populações estudadas, nenhuma diferença estatística (P>0,05) foi observada entre as contagens encontradas nas amostras de leite coletadas nas fazendas (tanques de resfriamento) e do LC. Portanto, o gênero Pseudomonas spp. e a espécie P. fluorescens não foram os psicrotróficos mais frequentes nesta linha de transporte de leite estudada.

Lipolytic capacity of Pseudomonas spp. isolated from refrigerated raw milk / Capacidade lipolítica de Pseudomonas spp. isoladas de leite cru refrigerado

The psychrotrophic bacteria growth in refrigerated milk is responsible for the production of heat resistant enzymes, including those which have lipolytic capacity. Lipases can cause rancidity and off-flavor in pasteurized and UHT milk, milk powder and cheese. Pseudomonas spp. is the major genera responsible by lipases synthesis, mainly Pseudomonas fluorescens. The lipolytic capacity of Pseudomonas spp. (1182 strains) and P. fluorescens (158 strains) isolated from cooling tank and bulk milk transportation were tested. Pseudomonas spp. strains were isolated by Pseudomonas Agar Base, adding the supplement cephalotin, fusidic acid, cetrimide- CFC (30º C/48 h) and Pseudomonas Cetrimide Agar with 10% of glicerol (21º C/ 48 h) to determine P. fluorescens. The strains lipolytic capacity were tested with Tributyrin Agar (21 oC/72 h). Lipolytic indexes of Pseudomonas spp. were 51.4% and 67.2% in cooling tank and bulk milk transportation (p< 0.05), respectively. P. fluorescens lipolytic capacity indexes were 26.4% from strains isolated from cooling tank and 41% (p < 0.05) from strains isolated from bulk milk transportation. High lipolytic indexes had been found in the strains isolated from refrigerated raw milk sent to manufacturing. Refrigeration temperature and storage time may contribute to the difference observed between two points of collection evaluated in this study.

A multiplicação de bactérias psicrotróficas em leite refrigerado pode promover a síntese de enzimas resistentes ao calor, entre elas aquelas que tem capacidade lipolítica. As lipases podem promover rancidez, e defeitos de sabor em leite pasteurizado, UHT, leite em pós e queijos. Pseudomonas spp. é o principal gênero responsável pela síntese desta enzimas, principalmente a espécie Pseudomonas fluorescens. Avaliou-se a capacidade lipolítica de 1182 cepas de Pseudomonas spp. e 158 cepas de P. fluorescens isoladas de amostras de leite de tanques de resfriamento (TR) e caminhão tanque (CT) enviado ao beneficiamento. Foi utilizado o Agar base Pseudomonas com suplemento cephalotin, fusidic acid, cetrimide - CFC (30º C/48 h ) para isolamento das pseudomonas e Pseudomonas Cetrimide Agar com 10% de glicerol (21º C/ 48 h) para a espécie P. fluorescens. Para avaliação da capacidade lipolítica as cepas fora semeadas em Ágar Tributirina (21 ºC/72 h). O índice de lipólise das cepas de Pseudomonas spp. provenientes do leite do tanque de resfriamento e do caminhão tanque (p<0,05) foram de 51,4% e 62,7%, respectivamente. Foram também observados índices de capacidade lipolítica de P. fluorescens de 26,4% e 41% (p<0,05) para TR e CT, respectivamente. Altos índices de cepas lipolíticas foram encontrados nas cepas isoladas do leite cru refrigerado enviado ao beneficiamento. Temperatura de refrigeração e tempo de estocagem podem ter contribuído para a diferença observada na população lipolítica dos dois pontos de coleta avaliados.

Pseudomonas spp. and P. fluorescens: population in refrigerated raw milk / Pseudomonas spp. e P. fluorescens: população em leite cru refrigerado

Raw milk samples were collected from cooling tanks (after they cooled for 48 h) in five dairy farms and the corresponding bulk tank (bulk milk transportation, BMT) when they arrived to the industry. Routine physical chemical analyzes and quantification of psychrotrophic (Pseudomonas spp. and P. fluorescens) and aerobic mesophilic (AM) populations were performed. Only relative density and titratable acidity values for samples of milk from three farms were in agreement to the quality parameters required by law. In the BMT, only the protein content has not reached the minimum value established by law, and counting was performed for AM (>105 colony forming units (CFU) mL-1) and psychrotrophic bacteria (2.8x106CFU mL-1). Pseudomonas spp. counting corresponded to 17.9% of the psychrotrophic population, and P. fluorescens was 3.4% of Pseudomonas spp . count. In milk samples from dairy farms, counts were variable for AM (3.4x105 to 3.7 x107CFU mL-1), psychrotrophic (4.0x104 to 3.1x106CFU mL-1), Pseudomonas spp. (2.3x104 to 1.8x105CFU mL-1), and P. fluorescens (62 to 8.4x103CFU mL-1). For the populations studied, no statistical difference (P>0.05) was observed between counts reported in milk samples collected in dairy farms (cooling tanks) and BMT. Therefore, the genera Pseudomonas spp. and P. fluorescens were not the most frequent psychrotrophic bacteria in this studied milk transportation line.(AU)

Amostras de leite foram coletadas de tanques de refrigeração (após 48h de refrigeração) em cinco fazendas de produção e do respectivo caminhão tanque (leite de conjunto, LC) ao chegar à indústria beneficiadora. Análises físico químicas de rotina e quantificação das populações de bactérias psicrotróficas (Pseudomonas spp. e P. fluorescens) e aeróbios mesófilos (AM) foram realizadas. Apenas os valores de densidade relativa e acidez titulável de amostras de leite de três fazendas estavam de acordo com a legislação. No LC, apenas o teor de proteína não atingiu o valor mínimo estabelecido por lei, e a contagem foi realizada para AM (>105 unidades formadoras de colônias (UFC) mL-1) e psicrotróficos (2,8x106UFC mL-1). A contagem de Pseudomonas spp. correspondeu a 17,9% da população de psicrotróficos, e 3,4% da contagem de Pseudomonas spp. eram da espécie P. fluorescens . Nas amostras de leite das fazendas de produtoras, as contagens foram variáveis para AM (3,4x105a 3,7x107UFC mL-1), psicrotróficos (4,0x104 a 3,1x106UFC mL-1), Pseudomonas spp. (2,3x104 a 1,8x105UFC mL-1) e P. fluorescens (62 a 8,4x103UFC mL-1). Para as populações estudadas, nenhuma diferença estatística (P>0,05) foi observada entre as contagens encontradas nas amostras de leite coletadas nas fazendas (tanques de resfriamento) e do LC. Portanto, o gênero Pseudomonas spp. e a espécie P. fluorescens não foram os psicrotróficos mais frequentes nesta linha de transporte de leite estudada.(AU)

Pseudomonas spp. and P. fluorescens: population in refrigerated raw milk / Pseudomonas spp. e P. fluorescens: população em leite cru refrigerado

ABSTRACT: Raw milk samples were collected from cooling tanks (after they cooled for 48 h) in five dairy farms and the corresponding bulk tank (bulk milk transportation, BMT) when they arrived to the industry. Routine physical chemical analyzes and quantification of psychrotrophic ( Pseudomonas spp. and P. fluorescens ) and aerobic mesophilic (AM) populations were performed. Only relative density and titratable acidity values for samples of milk from three farms were in agreement to the quality parameters required by law. In the BMT, only the protein content has not reached the minimum value established by law, and counting was performed for AM (>105 colony forming units (CFU) mL-1) and psychrotrophic bacteria (2.8x106CFU mL-1). Pseudomonas spp. counting corresponded to 17.9% of the psychrotrophic population, and P. fluorescens was 3.4% of Pseudomonas spp . count. In milk samples from dairy farms, counts were variable for AM (3.4x105 to 3.7 x107CFU mL-1), psychrotrophic (4.0x104 to 3.1x106CFU mL-1), Pseudomonas spp. (2.3x104 to 1.8x105CFU mL-1), and P. fluorescens (62 to 8.4x103CFU mL-1). For the populations studied, no statistical difference (P>0.05) was observed between counts reported in milk samples collected in dairy farms (cooling tanks) and BMT. Therefore, the genera Pseudomonas spp. and P. fluorescens were not the most frequent psychrotrophic bacteria in this studied milk transportation line.

RESUMO: Amostras de leite foram coletadas de tanques de refrigeração (após 48h de refrigeração) em cinco fazendas de produção e do respectivo caminhão tanque (leite de conjunto, LC) ao chegar à indústria beneficiadora. Análises físico químicas de rotina e quantificação das populações de bactérias psicrotróficas ( Pseudomonas spp. e P. fluorescens ) e aeróbios mesófilos (AM) foram realizadas. Apenas os valores de densidade relativa e acidez titulável de amostras de leite de três fazendas estavam de acordo com a legislação. No LC, apenas o teor de proteína não atingiu o valor mínimo estabelecido por lei, e a contagem foi realizada para AM (>105 unidades formadoras de colônias (UFC) mL-1) e psicrotróficos (2,8x106UFC mL-1). A contagem de Pseudomonas spp. correspondeu a 17,9% da população de psicrotróficos, e 3,4% da contagem de Pseudomonas spp. eram da espécie P. fluorescens . Nas amostras de leite das fazendas de produtoras, as contagens foram variáveis para AM (3,4x105a 3,7x107UFC mL-1), psicrotróficos (4,0x104 a 3,1x106UFC mL-1), Pseudomonas spp. (2,3x104 a 1,8x105UFC mL-1) e P. fluorescens (62 a 8,4x103UFC mL-1). Para as populações estudadas, nenhuma diferença estatística (P>0,05) foi observada entre as contagens encontradas nas amostras de leite coletadas nas fazendas (tanques de resfriamento) e do LC. Portanto, o gênero Pseudomonas spp. e a espécie P. fluorescens não foram os psicrotróficos mais frequentes nesta linha de transporte de leite estudada.

Ovinocultura leiteira no Brasil: aspectos e fatores relacionados à composição, ao consumo e à legislação / Dairy sheep industry in Brazil: aspects and factors related to the composition, consumption and legislation

Ao longo dos últimos anos a ovinocultura leiteira vem ganhando espaço no mercado internacional e nacional brasileiro. Desta forma, esta revisão teve como objetivo projetar um delineamento da ovinocultura leiteira brasileira, com enfoque nos aspectos da legislação, da composição e da qualidade do leite. Sua exploração mostra-se bastante vantajosa e rentável, visto que, o leite de ovelha carrega consigo características tecnológicas importantes para a indústria de alimentos. Seu elevado potencial de ser transformado em derivados lácteos, que agregam alta qualidade nutricional, atrai investidores e consumidores. As altas concentrações de sólidos totais presentes no leite de ovelha permitem a formulação de queijos, iogurtes e manteigas, de rentabilidade superior a de outras espécies. Fatores como a raça, a alimentação, a estação do ano, o estágio de lactação e o estado sanitário são variáveis que influenciam diretamente na composição do leite de ovelha. No Brasil, ainda não existe legislação específica para controle da qualidade do leite de ovinos, o que demonstra a necessidade de sua criação. São poucos os estudos brasileiros que caracterizam a composição e qualidade do leite de ovelha produzido no Brasil, sendo assim, importante o estudo das suas características e fatores que determinam sua composição.

Over the past few years, the dairy sheep industry have been taking space in the Brazilian national and international market. In this way, this work aimed to design a delineation of the Brazilian dairy sheep industry, with focus on aspects of the legislation, the composition and quality of milk. The explore shows up quite advantageous and profitable, since the sheep's milk carries technological characteristics important for the food industry. Their high potential to be transformed into milk products, that add high nutritional quality, attracts investors and consumers. The high concentrations of total solids present in the sheep milk allow the formulation of cheeses, yogurts and butters, of higher profitability than other species. Factors like the breed, feed, the season, the stage of lactation and the health status are variables that directly influence on composition of sheep's milk. In Brazil, there is still no specific legislation to control the quality of sheep milk, which demonstrates the necessity of its creation. There are few Brazilian studies that characterize the composition and quality of sheep milk produced in Brazil, being important the study of its characteristics and factors that determine its composition.

Ovinocultura leiteira no Brasil: aspectos e fatores relacionados à composição, ao consumo e à legislação / Dairy sheep industry in Brazil: aspects and factors related to the composition, consumption and legislation

Ao longo dos últimos anos a ovinocultura leiteira vem ganhando espaço no mercado internacional e nacional brasileiro. Desta forma, esta revisão teve como objetivo projetar um delineamento da ovinocultura leiteira brasileira, com enfoque nos aspectos da legislação, da composição e da qualidade do leite. Sua exploração mostra-se bastante vantajosa e rentável, visto que, o leite de ovelha carrega consigo características tecnológicas importantes para a indústria de alimentos. Seu elevado potencial de ser transformado em derivados lácteos, que agregam alta qualidade nutricional, atrai investidores e consumidores. As altas concentrações de sólidos totais presentes no leite de ovelha permitem a formulação de queijos, iogurtes e manteigas, de rentabilidade superior a de outras espécies. Fatores como a raça, a alimentação, a estação do ano, o estágio de lactação e o estado sanitário são variáveis que influenciam diretamente na composição do leite de ovelha. No Brasil, ainda não existe legislação específica para controle da qualidade do leite de ovinos, o que demonstra a necessidade de sua criação. São poucos os estudos brasileiros que caracterizam a composição e qualidade do leite de ovelha produzido no Brasil, sendo assim, importante o estudo das suas características e fatores que determinam sua composição.(AU)

Over the past few years, the dairy sheep industry have been taking space in the Brazilian national and international market. In this way, this work aimed to design a delineation of the Brazilian dairy sheep industry, with focus on aspects of the legislation, the composition and quality of milk. The explore shows up quite advantageous and profitable, since the sheep's milk carries technological characteristics important for the food industry. Their high potential to be transformed into milk products, that add high nutritional quality, attracts investors and consumers. The high concentrations of total solids present in the sheep milk allow the formulation of cheeses, yogurts and butters, of higher profitability than other species. Factors like the breed, feed, the season, the stage of lactation and the health status are variables that directly influence on composition of sheep's milk. In Brazil, there is still no specific legislation to control the quality of sheep milk, which demonstrates the necessity of its creation. There are few Brazilian studies that characterize the composition and quality of sheep milk produced in Brazil, being important the study of its characteristics and factors that determine its composition.(AU)

Losses in milk production and quality due to milk somatic cell count and heat stress of Holsteins cows in temperate climate / Perdas na produção e qualidade do leite devido contagem de células somáticas no leite e estresse térmico de vacas da raça Holandesa em clima temperado

The aims of present study were to evaluate the effects of milk somatic cell count (SCC) and heat stress on yield and milk composition of cows in a herd for commercial production in a temperate region during the period 2008-2012. Data from the monthly milk test-day records of 161±9 Holstein, totaling 9,650 milkings, were provided by the Association of Holstein Cattle Breeders of Parana State, and analyzed by descriptive analysis, correlation, analysis of variance, and regression analysis. The average daily milk yield was 31.78 kg/cow, which decreased to 29.31% when the somatic cell score (SCS) was 9, and to 11% when the Equivalent Temperature Index (ETI) was 32 or above. Lactose content decreased from SCS 0 until 9 and fat content decreased from SCS 1 until 9, totaling decrease 7.88 and 9.23%, respectively, when the SCS was 9. An opposite effect was observed for the protein content, which increased by 3.6% at SCS 8, when compared to SCS 0. Losses were observed in the daily total solids production from the SCS 0, totaling 30.64% at SCS 9.The increase in ETI to 32 or above reduced all milk constituents as much as 3.42%, except protein. These results, combined with the losses in milk yield at that ETI level, led to a decrease of up to 12.74% of milk solids. It is concluded that since losses in milk quality and yield resulting from SCC and ETI are significant, actions to prevent infection in the mammary gland and to provide a comfortable environment for dairy cattle are needed even in temperate regions.(AU)

O objetivo deste estudo foi avaliar a influencia da contagem de células somáticas (CCS) no leite e o estresse térmico sobre a produção e composição do leite de vacas de um rebanho de produção comercial em região de clima temperado, durante o período de 2008 a 2012. Os dados do dia de controle leiteiro de 161±9 vacas Holandesas, totalizando 9650 ordenhas, foram fornecidos pela Associação Paranaense de Criadores de Bovinos da Raça Holandesa e analisados através de estatística descritiva, correlação, análise de variância e regressão. A produção média diária de leite foi de 31,78 kg/vaca, com diminuição de 29,31% quando o escore de contagem de células somáticas (ECS) foi 9 e até 11% quando o Índice de Temperatura Equivalente (ITE) foi 32 ou maior. Os teores de lactose decresceram a partir do ECS 0 até 9 e de gordura a partir de ECS 1 até 9, totalizando diminuição de 7,88 e 9,23%, respectivamente, quando o ECCS foi 9. Um efeito inverso foi observado em relação ao teor de proteína, o qual aumentou 3,6% com ECS 8, comparado com o ECS 0. A produção diária de sólidos totais iniciou perdas a partir do ECS 0 e totalizou perdas de 30,64% quando este foi 9. O aumento do ITE até 32 ou mais diminuiu a concentração de todos os componentes do leite, exceto de proteína, em valores de até 3,42% da concentração. Estes efeitos, somados a diminuição na produção diária de leite com este nível de ITE, totalizaram perdas de até 12,74% na produção de sólidos totais. Conclui-se que as perdas de produção e qualidade de leite com o aumento da CCS e ITE são significativas e podem justificar a adoção de medidas para prevenir infecções na glândula mamária e conforto de vacas leiteiras, mesmo em região de clima temperado.(AU)

Occurrence of pathogens causing subclinical mastitis in Jaguapitã county, state of Paraná - Brazil / Ocorrência de patógenos causadores de mastite subclínica no município de Jaguapitã, estado do Paraná - Brasil

O objetivo desta pesquisa foi identificar a ocorrência dos patógenos causadores de mastite subclínica em um rebanho leiteiro tipo B no município de Jaguapitã, estado do Paraná, Brasil. Foram realizados 400 testes de Califórnia Mastite Teste (CMT) em amostras de leite de 100 animais, totalizando 400 tetos. Dentre os animais testados 55% reagiram ao CMT apresentando grau dois ou superior, com 157 tetos positivos. Após as amostras de leite dos 157 tetos serem submetidos à cultura em ágar sangue, 25,48% (40/157) não apresentaram crescimento ou houve crescimento de mais de duas colônias bacterianas, 28,03% (44/157) foram observadas Staphylococcus coagulase negativa (CNS), 8,28% (13/157) Streptococcus uberis, 7,64% (12/157) Staphylococcus aureus, 7,64% (12/157) Corynebacterium spp, 7,01% (11/157) Staphylococcus intermedius, 4,46% (7/157) Staphylococcus hyicus, 3,82% (6/157) Bacillus spp., 2,55% (4/157) para Streptococcus dysgalacteae, Enterobactéria e Leveduras. Conclui-se que a CNS é o mais relevante agente causador de mastite subclínica.(AU)

The aim of this research was to identify the occurrence of pathogens causing subclinical mastitis in grade B milk farms of the Jaguapitã county, state of Paraná, Brazil. California Mastitis Test (CMT) were carried out in 400 milk samples from 100 animals and 157 teats from 55 animals (55%) were positive, showed score two or higher to CMT. When these 157 positive samples to CMT were transported for bacterial culture in blood agar, 25.48% (40/157) samples showed no bacterial growth or more than two types of bacterial colonies grew, 28.03% (44/157) were Coagulase-negative staphylococci (CNS), 8.28% (13/157) were Streptococcus uberis, 7.64% (12/157) were Staphylococcus aureus, 7.64% (12/157) were Corynebacterium spp, 7.01% (11/157) were Staphylococcus intermedius, 4.46% (7/157) were Staphylococcus hyicus, 3.82% (6/157) were Bacillus spp., 2.55% (4/157) were Streptococcus dysgalacteae, Enterobacteria and Yeasts. We conclude that CNS is the most relevant subclinical mastitis causative agent.(AU)

Relationship between shape of teat and teat tip and somatic cell count (SCC) in dairy cows

Background: Mastitis is characterized by inflammation of the mammary gland, usually caused by pathogens, these infections may be classified as either clinical or subclinical, which are responsible for physical, chemical and bacteriological changes in milk and/or changes in the glandular tissue. The shape of both teat and teat tip are among the factors that may predispose to the development of clinical and subclinical mastitis, it is therefore important that they have a desirable morphology so that they are less susceptible to pathogens. The aim of this study was to investigate the relationship between teat shape and Somatic Cell Count (SCC) in dairy cattle.Materials, Methods & Results: The research was carried out in two dairy farms located in the municipality of Arapongas, Parana state, Brazil. Data were obtained from 150 Holstein cows variety black and white, where 597 teat shape and teat tip were evaluated during entry into the milking parlor. The teat shape was classified as desirable; bottle-shaped; cone-shaped; short; funnel-shaped; plump; and pencil-shaped, while the teat tip was classified as round; flat; funnel shape (inverted); disk, and pointed (hyperkeratosis). The somatic cell count (SCC) was carried out by flow cytometry by the equipment Somacount 500®1. The variable SCC was evaluated taking into account the shape of the teat, then two groups were formed...(AU)

Sheep milk: physical-chemical characteristics and microbiological quality.

Sheep milk is the third most consumed milk in Brazil. It is much appreciated for its nutritional status and is important for children that have problems with cow milk. Little information is known about the chemical, physical and microbiological composition of sheep milk from South Brazil. Thus, the aim of this study was to describe chemical and microbiological characteristics of sheep milk produced on two rural properties located in southern Brazil (ParanA and Rio Grande do Sul). The chemical composition of sheep milk was 17.32 g/100 g total solids, 5.86 g/100 g total protein, 4.46 g/100 g casein, 1.08 g/100 g whey protein, 7.28 g/100 g fat, 0.93 g/100 g ash, and 3.41 g/100 g lactose. High somatic cell count (1.7x106 cells/mL), total mesophilic bacterias (16.0 x 106 CFU/mL) and psychrotrophics (5.8 x 106 CFU/mL) were observed. Growth of Staphylococcus aureus, enterobacteria and coliforms occurred in 100% of the samples, and 45% of the samples showed growth of Escherichia coli. The sheep milk physical-chemical and microbiology parameters are similar to those presented in the literature for other countries but somatic cell count presented high levels.

Occurrence of pathogens causing subclinical mastitis in Jaguapitã county, state of Paraná - Brazil / Ocorrência de patógenos causadores de mastite subclínica no município de Jaguapitã, estado do Paraná - Brasil

O objetivo desta pesquisa foi identificar a ocorrência dos patógenos causadores de mastite subclínica em um rebanho leiteiro tipo B no município de Jaguapitã, estado do Paraná, Brasil. Foram realizados 400 testes de Califórnia Mastite Teste (CMT) em amostras de leite de 100 animais, totalizando 400 tetos. Dentre os animais testados 55% reagiram ao CMT apresentando grau dois ou superior, com 157 tetos positivos. Após as amostras de leite dos 157 tetos serem submetidos à cultura em ágar sangue, 25,48% (40/157) não apresentaram crescimento ou houve crescimento de mais de duas colônias bacterianas, 28,03% (44/157) foram observadas Staphylococcus coagulase negativa (CNS), 8,28% (13/157) Streptococcus uberis, 7,64% (12/157) Staphylococcus aureus, 7,64% (12/157) Corynebacterium spp, 7,01% (11/157) Staphylococcus intermedius, 4,46% (7/157) Staphylococcus hyicus, 3,82% (6/157) Bacillus spp., 2,55% (4/157) para Streptococcus dysgalacteae, Enterobactéria e Leveduras. Conclui-se que a CNS é o mais relevante agente causador de mastite subclínica.

The aim of this research was to identify the occurrence of pathogens causing subclinical mastitis in grade B milk farms of the Jaguapitã county, state of Paraná, Brazil. California Mastitis Test (CMT) were carried out in 400 milk samples from 100 animals and 157 teats from 55 animals (55%) were positive, showed score two or higher to CMT. When these 157 positive samples to CMT were transported for bacterial culture in blood agar, 25.48% (40/157) samples showed no bacterial growth or more than two types of bacterial colonies grew, 28.03% (44/157) were Coagulase-negative staphylococci (CNS), 8.28% (13/157) were Streptococcus uberis, 7.64% (12/157) were Staphylococcus aureus, 7.64% (12/157) were Corynebacterium spp, 7.01% (11/157) were Staphylococcus intermedius, 4.46% (7/157) were Staphylococcus hyicus, 3.82% (6/157) were Bacillus spp., 2.55% (4/157) were Streptococcus dysgalacteae, Enterobacteria and Yeasts. We conclude that CNS is the most relevant subclinical mastitis causative agent.

Losses in milk production and quality due to milk somatic cell count and heat stress of Holsteins cows in temperate climate / Perdas na produção e qualidade do leite devido contagem de células somáticas no leite e estresse térmico de vacas da raça Holandesa em clima temperado

The aims of present study were to evaluate the effects of milk somatic cell count (SCC) and heat stress on yield and milk composition of cows in a herd for commercial production in a temperate region during the period 2008-2012. Data from the monthly milk test-day records of 161±9 Holstein, totaling 9,650 milkings, were provided by the Association of Holstein Cattle Breeders of Parana State, and analyzed by descriptive analysis, correlation, analysis of variance, and regression analysis. The average daily milk yield was 31.78 kg/cow, which decreased to 29.31% when the somatic cell score (SCS) was 9, and to 11% when the Equivalent Temperature Index (ETI) was 32 or above. Lactose content decreased from SCS 0 until 9 and fat content decreased from SCS 1 until 9, totaling decrease 7.88 and 9.23%, respectively, when the SCS was 9. An opposite effect was observed for the protein content, which increased by 3.6% at SCS 8, when compared to SCS 0. Losses were observed in the daily total solids production from the SCS 0, totaling 30.64% at SCS 9.The increase in ETI to 32 or above reduced all milk constituents as much as 3.42%, except protein. These results, combined with the losses in milk yield at that ETI level, led to a decrease of up to 12.74% of milk solids. It is concluded that since losses in milk quality and yield resulting from SCC and ETI are significant, actions to prevent infection in the mammary gland and to provide a comfortable environment for dairy cattle are needed even in temperate regions.

O objetivo deste estudo foi avaliar a influencia da contagem de células somáticas (CCS) no leite e o estresse térmico sobre a produção e composição do leite de vacas de um rebanho de produção comercial em região de clima temperado, durante o período de 2008 a 2012. Os dados do dia de controle leiteiro de 161±9 vacas Holandesas, totalizando 9650 ordenhas, foram fornecidos pela Associação Paranaense de Criadores de Bovinos da Raça Holandesa e analisados através de estatística descritiva, correlação, análise de variância e regressão. A produção média diária de leite foi de 31,78 kg/vaca, com diminuição de 29,31% quando o escore de contagem de células somáticas (ECS) foi 9 e até 11% quando o Índice de Temperatura Equivalente (ITE) foi 32 ou maior. Os teores de lactose decresceram a partir do ECS 0 até 9 e de gordura a partir de ECS 1 até 9, totalizando diminuição de 7,88 e 9,23%, respectivamente, quando o ECCS foi 9. Um efeito inverso foi observado em relação ao teor de proteína, o qual aumentou 3,6% com ECS 8, comparado com o ECS 0. A produção diária de sólidos totais iniciou perdas a partir do ECS 0 e totalizou perdas de 30,64% quando este foi 9. O aumento do ITE até 32 ou mais diminuiu a concentração de todos os componentes do leite, exceto de proteína, em valores de até 3,42% da concentração. Estes efeitos, somados a diminuição na produção diária de leite com este nível de ITE, totalizaram perdas de até 12,74% na produção de sólidos totais. Conclui-se que as perdas de produção e qualidade de leite com o aumento da CCS e ITE são significativas e podem justificar a adoção de medidas para prevenir infecções na glândula mamária e conforto de vacas leiteiras, mesmo em região de clima temperado.