Parasites in dairy cattle farms in southern Brazil

Objective. The aim of this study was to investigate the relation among type and size of the farms related to the management of production and especially the presence and control of parasites. Materials and methods. To accomplish the above, 65 farms were used: 56 of semi-extensive system (cows in pastures), classified as low density farms (n=15, up to 15/cows), medium density farms (n=20; between 16-30/cows) and large density farms (n=21; over 31/cows). It was also included nine farms freestall system (n=9; confinement, over 70 cows) as part of the study population. Results. In the farms visited for the study can be noticed that the farms that had the higher level of technological improvement in managements practice were properties in high level (3) and freestall (4). In most of the farms there were observed the presence of ticks and flies, regardless of density of farms. Feces samples collected from 650 cows (n=10/farm), and analyzed using McMaster showed that 191 cows were positive for parasites. The presence of antibodies in bulk milk tank occurred in 36 (55.38%; CI95% 42-67) to Neospora caninum of the studied farms. Conclusions. We also conclude that N. caninum is present in cattle herds in Western Santa Catarina, and may be linked to reproductive problems in cows.


INTRODUCTION
The milk industry is an important component of agribusiness in Western Santa Catarina, Brazil.From this activity, the state reached an important contribution to gross domestic product (GDP) of the state of Santa Catarina, where dairy agribusiness still plays an extremely important role for the state, responsible for the survival of thousands of families in the countryside and in the generation of numerous direct and indirect jobs.Sanitary control herds are generally inefficient and can cause problems in production system, since they may be caused by various etiological agents, such as viruses, bacteria, fungi, and parasites (1,2).Helminths are among the diseases that most affect livestock productivity in many regions of the world.
In the 70s, it was estimated that each year about 10 million cattle and buffaloes died from consequences direct or indirect in presence of helminths (1).At present, it is believed that the number of animals with a fatal affection by parasites was reduced, due to strategic programs and new drugs; although significantly increased the diagnosis of treatmentresistant helminth (3).The damage caused by endoparasites can occur directly and indirectly reflected in weight gain, milk production, costs with antiparasitic, and mortality (1)(2)(3).
One of the main parasite that affects the Brazilian herd is the cattle tick Rhipicephalus (Boophilus) microplus, an ectoparasite that causes important economic losses in domestic cattle.Because of its ease way of reproduction, causes discomfort to the animals, as well as affect the development of the herd and the production of meat and milk.Furthermore, it can transmit diseases such as babesiosis and anaplasmosis, which can occur simultaneously and thus referred to as cattle tick fever (3).Besides the damage caused directly to the animal, the tick causes damage to the producer due to indirect spending with acaridae, labor and equipment necessities for application of such products (4).Another problem caused by tick relates to the environmental impact caused by the indiscriminate use of chemotherapeutic products.Over time, the difficulties in controlling the parasites increased significantly due to increasing resistance to chemotherapeutics in your control and treatment, often uncontrollable.Similarly, have
In this study we provide information about the Neospora caninum, the etiological agent responsible for neosporosis in cattle mainly.Recent studies have shown that several species of domestic animals can be infected, such as sheep, horses and goats (3,6).The disease in cattle have been associated with clinical signs such as abortion, absorption, mummification or birth of weak calves (7), but usually the calves are born healthy, but infected, because the N. caninum can be transmitted congenitally.Horizontal transmission, i.e. ingestion of oocysts shed in the feces of the dog has been mentioned as a risk factor for disease (8).Ticks, helminths and protozoa are responsible for diseases that cause economic problems for producers of milk will globally due to treatment costs and reduction in milk production (often unnoticed).In Santa Catarina Western, where you have a large milk cows stock, researches are limited.Therefore, the aim of this study was to investigate the relation between type and size of farms with production management and especially the presence of parasites, beyond the identification of risk factors related to infection by N. caninum.

MATERIAL AND METHODS
Local: Investigated properties.In this study 65 properties classified as specialized or semi specialized were surveyed (9) sampled from 20 municipalities in the western of Santa Catarina, Brazil (Figure 1).Of these, 56 semi-extensive system (cows in pastures), classified in this study as small (Class 1: n=15; up to 15 cows), medium (Class 2: n=20; between 16 and 30 cows) and large (Class 3: n=21; over 31 cows).Also as part of the study nine properties freestall system (Class 4: n=9; confinement, over 70 cows).The survey was conducted between November of 2013 and February of 2014, being applied a questionnaire for data collection, in order to verify the characteristics of the properties, as well as production and health management (Table 1).

Sample collection.
During the visit at the farm was performed of collection of feces of 10 lactating cows, as well as a bulk milk tank.Feces were collected directly from the rectum, allocated into plastic containers, identified and stored in insulated cooler bags until analysis in the laboratory.
Parasitological analysis of feces.In the laboratory, the feces were processed by the problema causado por las garrapatas se relaciona con el impacto ambiental generado por el uso indiscriminado de productos quimioterápicos.Con el tiempo, las dificultades en el control de los parásitos se incrementaron debido al aumento de la resistencia a la quimioterapia en su control y tratamiento, que a menudo resulta incontrolable.Asimismo, se han producido con el control de moscas, entre estos destaca la mosca domestica (a menudo relacionada con mastitis) y Haematobia irritans, un insecto hematófago que ha causado graves daños a la ganadería (3,5).
Las garrapatas, helmintos y protozoos son responsables de enfermedades que causan problemas económicos a los productores de leche a nivel mundial debido a los costos de tratamiento y la reducción en la producción de leche (a menudo inadvertida).En Santa Catarina occidental, donde hay un gran número de vacas lecheras, las investigaciones son limitadas.Por lo tanto, el objetivo de este estudio fue investigar la relación entre el tipo y el tamaño de las fincas con gestión de la producción y especialmente la presencia de parásitos, más allá de la identificación de los factores de riesgo relacionados con la infección por N. caninum.
Serologic assay for Neospora caninum in bulk milk tank.Milk collected from the tank of each farm was centrifuged at 7000 g for 10 minutes to obtain milk serum.So, the presence of antibodies to N. caninum in milk was verified by indirect immunofluorescence assay (IFA) using blades made with the protozoan tachyzoites (10).The samples considered positive were those in which full fluorescence from tachyzoites of N caninum was observed, as described by Camillo et al. (11).To validate the tests, samples known as positive and negative were used as reaction control.
Data analysis.The relationship between the size of the properties for reproductive management and presence of parasites was performed only in a descriptive manner, seeking to identify management practices and sanitary control measurements of every classified type of farm.para la recolección de datos con el fin de verificar las características de los predios, así como la gestión de la producción y la salud (Tabla 1).Recolección de muestras.La recolección de heces de 10 vacas así como un tanque de leche a granel se realizó durante la visita a las fincas.Las heces fueron recogidas directamente del recto, asignadas a envases plásticos, identificadas y almacenadas en bolsas frías aisladas hasta su análisis en el laboratorio.
Entre las fincas investigadas, tan solo el 9.23% tuvieron un resultado negativo para parásitos en heces, es decir, ninguna de las vacas mostró huevos de helmintos y ooquistes de coccidios en las heces.Entre los animales que tuvieron unos resultados Pascoeti -Parasites in dairy cattle farms in southern Brazil feces.Among animals positive for helminths, was observed variation between cows 50-3650 EPG, but in general, and average of 25.6 EPG/animal.However, 80% of animals had positive EPG less than 300.Among the animals positive for coccidia, was detected variation between cows 50-3050 OOPG, but the overall average was 16.7 OOPG/animal.
No significant association was found between EPG counts and (farm class production; Helminthes control and OOPG count), as well as no significant association was found between OOPG counts and (farm class production; Helminthes control and EPG count).The oocysts in the faeces were identified as the genus Eimeria.The helminth eggs identified in the feces of cattle were of Trichostrongylus spp.(35.28%),Haemonchus spp.(6.15%), Teladorsagia spp.(1.53%), Oesophagostomun spp.
(1.53%).In 33.84% of the farms was found mixed infection by two or three helminths.

Antibodies for N. caninum in milk.
The antibodies in milk farms were detected in 36 farms (55%; CI 95 % 42-67) (Figure 1).When we consider the classification of the farm, was found higher occurrence in farms from class 1 (Table 2).In this study, a significant association was observed between following factors: classification of the farms (small, medium, large; freestall); providing concentrated to the cattle and access of dogs to the local storage of inputs supplied to animals with positive farms for neosporosis.Medium and large farms had higher risks of infection by N. caninum (PR=5.86-IC95% :

DISCUSSION
The classification of the farm by number of dairy cows provided to verify numerical difference regarding managements, production and sanitary control, something expected due to the different degrees of technological improvement.For example, levels of protein and fat in milk are smaller farms that have a higher level of production technology, but in return these farms have lower number of somatic cell count (SCC) and total bacterial count (TBC) (9).The explanation for this behavior may be due to improved nutrition of animals and use of specialized breeds with the increase in production levels.Another point related to the production this hygiene practices at farm, observed in this study in a good sizeable percentage (pre-dipping, post-dipping and mastitis test); however, these practices were lower on farms of small and medium size (class 1 and 2) the extensive system.According of Simioni et al ( 9) and Martins et al (15), in specialized farms, the dairy cattle shows greater economic importance, encouraging the farmers to adopt appropriate hygiene practices during milking, and reproduction of the herd; similar to that observed in this study.
Ectoparasites are also factors that can cause loss in milk production, and the farms visited the main ectoparasites were identified to flies (Musca domestica and Haematobia irritans) and ticks (Rhipicephalus microplus).These flies can cause chronic dermatitis in cattle (horn fly), as well as being vectors of mastitiscausing bacteria (16), and nematode that causes stefanofilariosis, characterized by skin lesions, cranial to the mammary gland of the cow, caused by the genus Stephanofilaria (17), observed in this study, in a high percentage (56%) farms investigated.Approximately 62% of farms are facing serious problems with outbreaks of flies during the year, independent of the class 1 to 4 in this study.The presence of composters can help control flies (3), but only 60% used on farms in the west of Santa Catarina State.
According to the literature, ticks are the most important ectoparasites for dairy farming, and approximately 75% of the cattle population is affected by this parasite.Tropical and subtropical regions are favorable for the development of ticks ( 4), target region this study.In addition to direct damage to the animal, the tick transmits Babesia spp.and Anaplasma spp.(3), responsible for causing one of the most common diseases (bovine parasitic sadness) in dairy cows in the Brazilian deberse a la mejora en la nutrición de los animales y el uso de razas especializadas con el aumento correspondiente en los niveles de producción.Otro punto relacionado con la producción es prácticas higiénicas en la finca, observadas en este estudio en un porcentaje considerable (prueba de inmersión previa, inmersión posterior y mastitis); sin embargo, estas prácticas fueron inferiores en granjas de pequeño y mediano tamaño (clase 1 y 2) del sistema extensivo.Según Simioni et al ( 9) y Martins et al (15), en granjas especializadas, el ganado lechero muestra una mayor importancia económica, alentando a los agricultores a adoptar prácticas de higiene adecuadas durante el ordeño y la reproducción de la manada; similar a lo observado en este estudio.
Aproximadamente el 62% de las granjas se enfrentan a graves problemas de brotes de moscas durante el año, independiente de la clase 1 a 4 en este estudio.La presencia de compostadores puede ayudar en el control de las moscas (3), pero son utilizados por tan solo el 60% de las granjas en el oeste del estado de Santa Catarina.
Según la literatura, las garrapatas son los ectoparásitos más importantes para la ganadería lechera, y aproximadamente el 75% de la población de ganado es afectada por este parásito.Las regiones tropicales y subtropicales resultan favorables para el desarrollo de las garrapatas (4), las cuales corresponden a la región objetivo de este estudio.Además del daño directo para el animal, las garrapatas transmiten Babesia spp y Anaplasma spp (3), los cuales son responsables de causar una de las enfermedades más comunes (tristeza parasitaria bovina) en vacas lecheras en los estados brasileños.En este estudio esto no fue diferente, se encontró que el 58% de las fincas tienen casos anuales de babesiosis y anaplasmosis, independientemente de la clasificación dada en el presente estudio.La enfermedad es una de las mayores pérdidas económicas debido a problemas sanitarios en el ganado debido a los costos del tratamiento, las altas tasas de mortalidad y morbilidad, así como reducción en la producción de states.This study was no different, as the interview was found that 58% of farms have annual cases of babesiosis and anaplasmosis, regardless the classification given in this study.
The disease is a sanitary problems greatest economic loss in cattle due to treatment costs, high rates of mortality and morbidity, as well as reduction in the production of meat and/or milk (3,18).Most farms investigated make use of antiparasitic to control ticks, and only 16% of farms do not tick control, because according to the producers the degree of infestation is low.This can be explained because these farms are rotational grazing system or animals are in confinement (freestall), which reduces the chance of development of the parasite, and contact with animals. The .
Nuestro modelo fue capaz de identificar la presencia de perros como un factor de riesgo de N. caninum, este hallazgo ha sido ampliamente reportado en muchos estudios y es un factor epidemiológico muy importante puesto que los perros se convierten en anfitriones definitivos de N. caninum (20), pero lo que encontramos brinda más detalles en esta cuestión.Se encontró la posibilidad de que un perro tenga contacto con el alimento para ganado, lo que significa que los alimentos almacenados representan un difícil factor de riesgo para N. caninum infeccioso.Nuestros hallazgos son consistentes con otros que han demostrado que los perros de granja pueden defecar en los corredores de alimentación y en hierba almacenada o ensilado de maíz, lo que puede causar evidencia de infección puerperal bovina que los rebaños con ninguna evidencia de este tipo (21,22).Con base en estos resultados, puede estar justificado suponer que la contaminación de la zona de alimentación se encuentra más estrechamente vinculada con la infección por N. caninum (23).De manera complementaria al factor anterior, se encontró también que las granjas que ofrecen concentrado a los ganados eran más propensas a tener resultados positivos para N. caninum, este conocimiento es consistente con Topazio et al (6) donde se ha demostrado que las cabras de granja que con una rica dieta qué incluye concentrado tienen más Pascoeti -Parasites in dairy cattle farms in southern Brazil of the infections, and a possible explanation for these may due to inadequate food storage what may make possible a direct contact between fed and infected dogs.Finally, larger farms were identify on the study as class 2 and 3 were associated to risk of N. caninum infection, in a study from Italy, the risk of individual cattle becoming seropositive increased with the size of the herd.When the analysis was restricted to data from northern Italy, the number of dogs per farm interacted significantly with herd size; i.e., the risk of being seropositive increased in larger herds with an increasing number of dogs per farm (24) in addition a study conducted in Germany, larger herds had an increased risk of being bulk milk positive ( 25).An explanation for herd size as a risk factor could be that hygienic measures to prevent dogs from feeding on placentas or other infectious material are more difficult to follow with large herds than with small herds (25).
In summary, we can conclude that there are technical differences between the farms, which is reflected in handling and production, i.e. the largest farms (Class 3 and 4) can produce a greater volume of milk, as is best compensated, reflecting to increased investment in the farm (technification).The contamination by gastrointestinal parasites had no difference between production systems.Also concluded that over 50% of farms in western Santa Catarina has infected cows by N. caninum, which may be related to reproductive problems reported by producers.The detection of antibodies to N. caninum in collective samples of milk proved to be a great diagnostic tool to herd, reducing costs with individual tests in a research first.
Ethics committee.Experimental protocol was approved by the Animal Welfare Committee of Universidade do Estado de Santa Catarina (UDESC) under number 1.23.14.

Figure 1 .
Figure 1.Map of epidemiological distribution of positive samples derived from milk from the farm tank, i.e.collective samples (n=36) in a total of 65samples in Santa Catarina state, Brazil.

Table 1 .
Information on management practices and occurrence of some parasitic diseases in dairy cattle farm in western Santa Catarina State, Brazil.
25tibodies to Neospora caninum in milk.The data generated by the questioner applied records and by the test were recorded and analyzed in statistical software R, version 3.1 (R core team, 2012) and map was produced using Arc Map ® version 3.2.1 (ESRI, Redlands, CA, USA).Cross tabulation and descriptive statistics were performed.Variables were first screened based on the response variable (positive or negative for Neosporosis), variables with large amounts of missing data (>10%) and limited variability (<20%) were not included in the multivariable model.The remaining variables were entered individually into a univariable logistic regression model and selected for inclusion in the multivariable model if p<0.25.Subsequently, all the screened variables were submitted to correlation analysis.The non-collinear variable was <0.7.Interactions * Mean ± standard deviation; # Testing black mug.& Stephanofilariasis.+ Observation made on the day of collection, the researchers.

Table 2 .
Univariate analysis of risk factors forNeospora caninum infection in bulk milk tank from 65 farms, in the state of Santa Catarina, Brazil.Helminthes control and EPG count) and variability of the model was expressed by R 2 .
(14)resented as descriptive measurement is the mean and standard deviation.All variables were tested for normality of variance by Shapiro-Wilk test(13), skewness, kurtosis and homogeneity by Levene's test(14)previously to analysis by linear regression.Wald statistics was used to assess the significance of association (farm class production;

table 1 .
Considering the average among the surveyed properties without considering size and production system, it was found that the time working in the dairy business was 17.7 years, as well as the frequency of the total number of cows and lactating cows were 53.3 and 38.2%, respectively.The average milk production of the surveyed properties was 18.2 liters/day.From the properties it was found that the labor used in dairy cattle as familiar was 84.6%, outsourced (6.15%) and mixed (9.23% -family and outsourced).In percentages, other important data relating to the farms studied were: receivable amount (83.7%) and milk quality (85.4%), performing pre-dipping (70.8%), and post-dipping (88.8%), rapid test for mastitis (66.3%), reproductive control (95%), use of artificial insemination (86.2%), use of sexed semen (18.4%), providing commercial feed (86%) and silage (91.2%).Parasitological of faces.The results of EPG and OOPG (oocyst per gram) according to classification of the farms were presented in table 1. Between farms investigated, only 9.23% were negative for parasites in feces, i.e., none of the cows showed helminthes eggs and coccidia oocysts in the

Table 3 .
Multivariate analysis of risk factors forNeospora caninum infection in bulk milk tank from 65 farms, in the state of Santa Catarina, Brazil.