IgG and IgG2 antibodies from cattle naturally infected with Anaplasma marginale recognize the recombinant vaccine candidate antigens VirB9, VirB10, and elongation factor-Tu

Anaplasma marginale is an important vector-borne rickettsia of ruminants in tropical and subtropical regions of the world. Immunization with purified outer membranes of this organism induces protection against acute anaplasmosis. Previous studies, with proteomic and genomic approach identified 21 proteins within the outer membrane immunogen in addition to previously characterized major surface protein1a-5 (MSP1a-5). Among the newly described proteins were VirB9, VirB10, and elongation factor-Tu (EF-Tu). VirB9, VirB10 are considered part of the type IV secretion system (TFSS), which mediates secretion or cell-to-cell transfer of macromolecules, proteins, or DNA-protein complexes in Gram-negative bacteria. EF-Tu can be located in the bacterial surface, mediating bacterial attachment to host cells, or in the bacterial cytoplasm for protein synthesis. However, the roles of VirB9, VirB10, and TFSS in A. marginale have not been defined. VirB9, VirB10, and EF-Tu have not been explored as vaccine antigens. In this study, we demonstrate that sera of cattle infected with A. marginale, with homologous or heterologous isolates recognize recombinant VirB9, VirB10, and EF-Tu. IgG2 from naturally infected cattle also reacts with these proteins. Recognition of epitopes by total IgG and by IgG2 from infected cattle with A. marginale support the inclusion of these proteins in recombinant vaccines against this rickettsia.

Spoligotyping and variable number tandem repeat analysis of Mycobacterium bovis isolates from cattle in Brazil

We performed spoligotyping and 12-mycobacterial interspersed repetitive unit-variable number tandem repeats (MIRU-VNTRs) typing to characterise Mycobacterium bovis isolates collected from tissue samples of bovines with lesions suggestive for tuberculosis during slaughter inspection procedures in abattoirs in Brazil. High-quality genotypes were obtained with both procedures for 61 isolates that were obtained from 185 bovine tissue samples and all of these isolates were identified as M. bovis by conventional identification procedures. On the basis of the spoligotyping, 53 isolates were grouped into nine clusters and the remaining eight isolates were unique types, resulting in 17 spoligotypes. The majority of the Brazilian M. bovis isolates displayed spoligotype patterns that have been previously observed in strains isolated from cattle in other countries. MIRU-VNTR typing produced 16 distinct genotypes, with 53 isolates forming eight of the groups, and individual isolates with unique VNTR profiles forming the remaining eight groups. The allelic diversity of each VNTR locus was calculated and only two of the 12-MIRU-VNTR loci presented scores with either a moderate (0.4, MIRU16) or high (0.6, MIRU26) discriminatory index (h). Both typing methods produced similar discriminatory indexes (spoligotyping h = 0.85; MIRU-VNTR h = 0.86) and the combination of the two methods increased the h value to 0.94, resulting in 29 distinct patterns. These results confirm that spoligotyping and VNTR analysis are valuable tools for studying the molecular epidemiology of M. bovis infections in Brazil.

Genotyping Mycobacterium bovis from cattle in the Central Pampas of Argentina: temporal and regional trends

Mycobacterium bovis is the causative agent of bovine tuberculosis (TB), a disease that affects approximately 5% of Argentinean cattle. Among the molecular methods for genotyping, the most convenient are spoligotyping and variable number of tandem repeats (VNTR). A total of 378 samples from bovines with visible lesions consistent with TB were collected at slaughterhouses in three provinces, yielding 265 M. bovis spoligotyped isolates, which were distributed into 35 spoligotypes. In addition, 197 isolates were also typed by the VNTR method and 54 combined VNTR types were detected. There were 24 clusters and 27 orphan types. When both typing methods were combined, 98 spoligotypes and VNTR types were observed with 27 clusters and 71 orphan types. By performing a meta-analysis with previous spoligotyping results, we identified regional and temporal trends in the population structure of M. bovis. For SB0140, the most predominant spoligotype in Argentina, the prevalence percentage remained high during different periods, varying from 25.5-57.8% (1994-2011). By contrast, the second and third most prevalent spoligotypes exhibited important fluctuations. This study shows that there has been an expansion in ancestral lineages as demonstrated by spoligotyping. However, exact tandem repeat typing suggests dynamic changes in the clonal population of this microorganism.

Should I go to court?: an assessment of the role of the Judiciary in disputes between cattle raisers and meatpackers in Brazil

The main objective of this perspective paper is to analyze the role of the judiciary in resolving conflicts between cattle raisers and meatpacking firms in Brazil. Looking at the transaction for the acquisition of cattle for slaughter in the state of Mato Grosso do Sul (the central-western region of Brazil), the analysis encompasses three steps. First, the authors describe the transaction pattern between cattle raisers and meatpacking firms, identifying a guarantee vacuum within the supply chain. Secondly, the authors present evidence that the guarantee vacuum may give rise to legal conflicts of non-payment, which are indeed prevalent in disputes that reach the courts. Finally, the role of the judiciary in resolving these conflicts is investigated. Results suggest that, on average, producers have little confidence in the legal system, indicating the potential function that informal mechanisms may play in the supply chain. The paper concludes by suggesting some implications for public and private strategies.

Soil phosphorus fractions in sandy soils amended with cattle manure for long periods

Phosphorus fractions were determined in soil samples from areas fertilized or not with farmyard cattle manure (FYM) and in samples of FYM used in the semi-arid region of Paraiba state, Brazil. Soil samples were taken from the 0-20; 20-40 and 40-60 cm layers of 18 cultivated areas, which, according to interviews with farmers, had been treated with 12 to 20 t ha-1 FYM annually, for the past 2 to 40 years. Soil samples were also collected from four unfertilized pasture areas as controls. Phosphorus in the soil samples was sequentially extracted with water (Pw), resin (Pres), NaHCO3 (Pi bic and Po bic), NaOH (Pi hid and Po hid), H2SO4 (Pacid) and, finally, by digestion with H2SO4/H2O2 (Presd). Nine FYM samples were extracted with water, resin, Mehlich-1, H2SO4, NaOH or digestion with H2SO4/H2O2, not sequentially, and the extracts analyzed for P. The sampled areas had homogeneous, sandy and P-deficient soils; increases in total soil P (Pt) above the mean value of the control areas (up to 274 mg kg-1 in the 0-20 cm layer of the most P-enriched samples) were therefore attributed to FYM applications, which was the only external P input in the region. Regression analysis was used to study the relationship between soil P fractions and Pt. The Pacid fraction, related to Ca-P forms, showed the greatest increases (p < 0.01) as a result of FYM applications, rising from 8.4 mg kg-1 in a non-fertilized sample to 43.8 mg kg-1 in the sample with the highest Pt content. The sum of Pw, Pres and Pi bic, considered as labile P, showed comparable increases with Pacid, while Pi hid showed the smallest increase due to FYM applications. Organic P forms also increased, more so the fraction Po hid, considered less labile, than the more labile one, Po bic. The residual P fraction was practically half of Pt, independently of the Pt value. Increases in labile P, Pacid and organic P were justified by the high average concentration of Pw (36 %), Pacid (34 %), and Po hid (30 %) in the FYM. Significant changes in the proportion of P forms among soil layers indicated the downward movement of P in organic forms.

Behavioral avoidance tests to evaluate effects of cattle slurry and dairy sludge application to soil¹

The application of organic wastes to agricultural soils is not risk-free and can affect soil invertebrates. Ecotoxicological tests based on the behavioral avoidance of earthworms and springtails were performed to evaluate effects of different fertilization strategies on soil quality and habitat function for soil organisms. These tests were performed in soils treated with: i) slurry and chemical fertilizers, according to the conventional fertilization management of the region, ii) conventional fertilization + sludge and iii) unfertilized reference soil. Both fertilization strategies contributed to soil acidity mitigation and caused no increase in soil heavy metal content. Avoidance test results showed no negative effects of these strategies on soil organisms, compared with the reference soil. However, results of the two fertilization managements differed: Springtails did not avoid soils fertilized with dairy sludge in any of the tested combinations. Earthworms avoided soils treated with sludge as of May 2004 (DS1), when compared with conventional fertilization. Possibly, the behavioral avoidance of earthworms is more sensitive to soil properties (other than texture, organic matter and heavy metal content) than springtails

ORGANIC PHOSPHORUS FRACTIONS IN SOIL FERTILIZED WITH CATTLE MANURE

Inorganic phosphorus (Pi) usually controls the P availability in tropical soils, but the contribution of organic P (Po) should not be neglected, mainly in systems with low P input or management systems that promote organic matter accumulation. The aims of this study were to evaluate the changes in the Po fractions over time in soil fertilized and not fertilized with cattle manure and to correlate Po forms with available P extracted by anion exchange resin. The experiment was carried out under field conditions, in a sandy-clay loam Haplustox. The experimental design was a 2 × 9 randomized complete block factorial design, in which the first factor was manure application (20 t ha-1) or absence, and the second the soil sampling times (3, 7, 14, 21, 28, 49, 70, 91, and 112 days) after manure incorporation. Labile, moderately labile and non-labile Po fractions were determined in the soil material of each sampling. Manure fertilization increased the Po levels in the moderately labile and non-labile fractions and the total organic P, but did not affect the Po fraction proportions in relation to total organic P. On average, 5.1 % of total Po was in the labile, 44.4 % in the moderately labile and 50.5 % in the non-labile fractions. Available P (resin P) was more affected by the manure soluble Pi rather than by the labile Po forms. The labile and non-labile Po fractions varied randomly with no defined trend in relation to the samplings; for this reason, the data did not fit any mathematical model.

Follicular dynamics in zebu cattle

The aim of this study was to investigate the follicular dynamics during estrous cycle in Gir breed (Bos indicus) cows. Follicular growth and atresia during estrous cycle were evaluated using a portable ultrasound device. Luteal activity was evaluated by serum progesterone levels. Cycles with two (6.67%), three (60.00%), four (26.67%) and five (6.67%) follicular waves were observed. There was no difference (P>0.05) in dominant or subordinate follicles growth or atresia rates among follicular waves. The maximum diameter of the ovulatory follicle was higher than the diameter of the other dominant follicles in cycles with four waves, and higher than the diameter of the second dominant follicle in cycles with three waves (P<0.05). There was no difference (P>0.05) in estrous cycle length (21.11±1.76 and 22.25±1.71 days) or progesterone levels during diestrous (4.48±1.45 and 5.08±1.40 ng/mL) between cycles with three or four waves. Follicular dynamics in Gir cattle is characterized by a higher incidence of cycles with three or four waves, associated with a low persistence of the dominant follicle.

Daily intake of lactating crossbred cows grazing elephant grass rotationally

The goal of this trial was to estimate the total dry matter (TDMI) and daily pasture dry matter intakes (PDMI) by lactating crossbred Holstein - Zebu cows grazing elephant grass (Pennisetum purpureum Schum.) paddocks submitted to different rest periods. Three groups of 24 cows were used during two years. The paddocks were grazed during three days at the stocking rate of 4.5 cows/ha. Treatments consisted of resting periods of 30 days without concentrate and resting periods of 30, 37.5 and 45 days with 2 kg/cow/day of 20.6% crude protein concentrate. From July to October, pasture was supplemented with chopped sugarcane plus 1% urea. Total daily dry matter intake was estimated using the extrusa in vitro dry matter digestibility and the fecal output with chromium oxide. Regardless of the treatment the estimated average TDMI was 2.7, 2.9 and 2.9±0.03% and the mean PDMI was 1.9, 2.1 and 2.1±0.03% of body weight in the first, second and third grazing day, respectively (P<0.05). Only during the summer pasture quality was the same whichever the grazing day. Sugarcane effectively replaced grazing pasture, mainly in the first day when pasture dry matter intake was lowest.

Larvicidal potential of Sapindus saponaria to control the cattle tick Boophilus microplus

The objective of this study was to evaluate the larvicidal potential of a crude ethanol extract (CEE) of soapberry Sapindus saponaria stem peel on the cattle tick Boophilus microplus. Tick larvae obtained by incubating engorged females, collected from naturally infested cattle, were placed in envelopes of filter paper impregnated with different concentrations of CEE in the test group, and distilled water in the control group. Four repetitions were made with each solution (n>120). Mortality was observed after 48 hours. Lethal concentration values of 1,258 ppm (LC50) and 6,360 ppm (LC99) were obtained.

Cytogenetic and molecular analysis of the Pantaneiro cattle breed

The objective of this work was to characterize Pantaneiro cattle genetically through its paternal ancestry by the morphology of the Y chromosome, whether submetacentric or acrocentric, as well as to identify the maternal ancestry through mitochondrial DNA. The karyotype and mitochondrial DNA of 12 bulls of Pantaneiro breed were analyzed. The Y chromosome was analyzed in lymphocyte metaphases and the mitochondrial DNA by diagnosing its haplotype (Bos taurus and Bos indicus). Among Pantaneiro animals analyzed three had a taurine (submetacentric) Y and nine had a zebuine (acrocentric) Y chromosome, suggesting breed contamination by Zebu cattle, once Pantaneiro is considered to be of European origin. The mitochondrial DNA was exclusively of taurine origin, indicating that the participation of zebuines in the formation of the breed occurred entirely through the paternal line.

Body condition score to predict the postpartum fertility of crossbred beef cows

The relationship between changes in body condition score (BCS) during the postpartum and fertility in beef cows suckling calves under extensive conditions were investigated. Cows were subjected to four BCS evaluations over the postpartum period, starting around one month after calving. In the second evaluation cows were treated with medroxy-progesterone acetate impregnated pessaries and received an injection of estradiol benzoate. At the third evaluation, pessaries were removed and calves were separated from the cows for 96 hours, during which time estrous was observed twice a day, and animals artificially inseminated 12 hours after detection. When calves returned to their dams, bulls were introduced until a 60-day mating period was reached. The distribution of BCS differed among calving groups and evaluations. Results indicated that only cows comprising a BCS 3 (1 to 5 scale) around the first month postpartum can be used in an artificial insemination program with possibilities of becoming pregnant. There was no statistical difference between the calving groups in pregnancy rate. The evolution of the BCS of the cows during postpartum can be used to adjust the start of the breeding season to coincide with the time of the year where herd pregnancy rates will be highest.

Tannin-tolerant bacteria from crossbred Holstein x Zebu cows

The objective of this work was to isolate and characterize tannin-tolerant ruminal bacteria from crossbred Holstein x Zebu cows fed a chopped mixture of elephant grass (Pennisetum purpureum), young stems of "angico-vermelho" (Parapiptadenia rigida), and banana tree (Musa sp.) leaves. A total of 117 bacteria strains were isolated from enrichment cultures of rumen microflora in medium containing tannin extracts. Of these, 11 isolates were able to tolerate up to 3 g L-1 of tannins. Classical characterization procedures indicated that different morphological and physiological groups were represented. Restriction fragments profiles using Alu1 and Taq1 of 1,450 bp PCR products from the 16S rRNA gene grouped the 11 isolates into types I to VI. Sequencing of 16S rRNA PCR products was used for identification. From the 11 strains studied, seven were not identifiable by the methods used in this work, two were strains of Butyrivibrio fibrisolvens, and two of Streptococcus bovis.

Factors associated with 56-day non-return rate in dairy cattle

The objective of this work was to identify factors associated with the 56-day non-return rate (56-NRR) in dairy herds in the Galician region, Spain, and to estimate it for individual Holstein bulls. The experiment was carried out in herds originated from North-West Spain, from September 2008 to August 2009. Data of the 76,440 first inseminations performed during this period were gathered. Candidate factors were tested for their association with the 56-NRR by using a logistic model (binomial). Afterwards, 37 sires with a minimum of 150 first performed inseminations were individually evaluated. Logistic models were also estimated for each bull, and predicted individual 56-NRR rate values were calculated as a solution for the model parameters. Logistic regression found four major factors associated with 56-NRR in lactating cows: age at insemination, days from calving to insemination, milk production level at the time of insemination, and herd size. First-service conception rate, when a particular sire was used, was higher for heifers (0.71) than for lactating cows (0.52). Non-return rates were highly variable among bulls. Asignificant part of the herd-level variation of 56-NRR of Holstein cattle seems attributable to the service sire. High correlation level between observed and predicted 56-NRR was found.

Hematological profile of beef cattle with divergent residual feed intake, following feed deprivation

The objective of this work was to characterize the hematological profiles of steers and bulls, according to residual feed intake (RFI), after feed deprivation. Twenty‑month‑old Nellore steers and bulls were fed feedlot diets for 70 and 56 days, respectively. RFI was calculated as the difference between actual feed intake and expected feed intake. More and less efficient steers and bulls, according to RFI, were subjected to 24 hours of food deprivation. Blood was sampled prior to and following the withdrawal period. Hematological analyses included total and differential white blood cell count, red blood cell count and morphology, and plasma glucose concentration. Variation in RFI did not influence plasma glucose concentration or blood cell profiles. Glucose concentrations in bulls decreased from 114 to 97 mg dL‑1, but remained unchanged in steers, and the neutrophil:lymphocyte ratio increased from 0.39 to 0.57 following deprivation. Hematological profiles do not differ between more and less efficient steers and bulls, according to RFI.