Comparison of duplex PCR and microscopic techniques for the identification of Babesia bigemina and Babesia bovis in engorged female ticks of Boophilus microplus

A single-step duplex polymerase chain reaction (PCR) technique and traditional microscopic examination of haemolymph smears were used to detect Babesia bigemina and/or Babesia bovis infection in engorged female ticks of Boophilus microplus recovered from calves raised in an endemic area of the State of Minas Gerais, Brazil. In the PCR amplification of tick-derived DNA, pairs of oligonucleotide primers specific for a 278-bp sequence from B. bigemina and for a 350-bp sequence from B. bovis were used conjointly. The microscopic examination of haemolymph revealed that 16.7% of the engorged ticks were infected with Babesia spp., although no significant differences (rho > 0.05) were found in the infection rate of ticks collected from calves of different age groups. PCR analysis showed that 77.8% of the engorged ticks whose haemolymph contained sporokinetes were infected with B. bigemina, 7.8% with B. bovis and 14.4% with both protozoan species. However, the PCR assay further revealed that, amongst the engorged female ticks whose haemolymph was apparently negative for the presence of sporokinetes, 15.6% were infected with B. bigemina, 2.2% with B. bovis and 10.0% with both species. The duplex PCR method is thus more efficient and sensitive than the microscopic assay and also permits facile identification of the protozoa species present in engorged female ticks.     

Brazilian foot and mouth disease status and meat exportation to the European Union

The aim of this study was to define the differences between the Brazilian states that export and do not export meat to the European Union (EU) and to identify the variables that are important to meet the export requirements. Infrastructure and computerization of the control of animal transit in Brazil that impact on regional health status were evaluated and linked to other variables such as status for foot and mouth disease (FMD) and qualification to export meat to EU. Variables related to transit control of bovines implemented by the state agencies of animal health and inspection in each Brazilian state were evaluated. Using a discriminant analysis, four variables were selected that explained the variation between Brazilian states that were “free” and “not free” of FMD while another four were selected to explain the variation between the zones “approved” and “not approved” to export meat to the EU, including number of official veterinarians, total transit of bovines and buffaloes, total number of animal transit certificates issued for bovine and buffaloes at the state or zone level, and total number of municipalities in the state or zone. It was possible to correctly discriminate between “free” and “not free” FMD states or zones. Variables related to animal transit are important in assessing the state for the classification of animal health situation and for EU approval for the exportation of meat.     

Short-wave solar radiation level willingly tolerated by lactating Holstein cows in an equatorial semi-arid environment

The amount of short-wave solar radiation willingly tolerated by lactating Holstein cows on pasture was determined in an equatorial semi-arid environment. The study was carried out on a dairy farm located in Limoeiro do Norte, CE, northeastern Brazil. The observed behaviours were as follows: grazing, under the sun, under the shade, standing, lying, ruminating, idling and wallowing in the water. The behaviours were recorded using instantaneous scan sampling at regular intervals of 15 min from 0600 to 1800 hours over 5 days. On all sampling days, the meteorological variables, including local short-wave solar radiation (R _S-W, W m^?2), were recorded. The R _S-W data were divided into five levels. The sun exposure was more frequent under low (100 %) and moderately low (97 %) levels, when R _S-W remained below 500 W m^?2. The grazing was more intense under low (100 %) and moderately low (93 %) levels. Above 500 W m^?2, the grazing time significantly decreased (11 %). The cows avoided grazing under high (0 %) and very high (0 %) levels, when R _S-W exceeded 700 W m^?2. The ruminating behaviour was more frequent under high (33 %) and very high (37 %) levels, in which the highest averages of R _S-W were recorded (815 and 958 W m^?2, respectively). The standing posture was more frequent under low (100 %) and moderately low (97 %) levels. Therefore, the critical R _S-W level that motivates cows to stop grazing and seek shade was in the interval between 500 and 700 W m^?2.     

Epidemiology of bovine anaplasmosis in dairy herds from Costa Rica

Bovine anaplasmosis is endemic and occurs in almost all areas of livestock production of Costa Rica. The aim of this study was to determine the seroprevalence and risk factors of anaplasmosis in dairy farms of Costa Rica by the recombinant truncated MSP-5 (rMSP-5) enzyme-linked immunosorbent assay (ELISA). Serum samples were obtained from 733 cattle from 20 commercial dairy herds of Costa Rica. The overall seroprevalence was 37.2% and herd seroprevalence ranged from 20.0 to 72.0%. The age-specific seroprevalence was 49.3% in young and 33.4% in adult animals. The main risk factors associated with seroprevalence were season of occurrence of clinical cases (rainy season) (OR=22.8), presence of tabanids (OR=9.5) and stable flies (OR=6.2), stable flies control measures (OR=3.2), non-use of ear tattoos (OR=2.8), interval of veterinary visit (≤ 60 days) (OR=2.7), altitude of the farms (<800 masl) (OR=2.6) and age (<2 years) (OR=1.8). The results indicated that exposure of cattle to Anaplasma marginale is common in dairy herds of Costa Rica and endemic instability situation probably is due to inadequate vector control.     

Exogenous paraoxonase‐1 during oocyte maturation improves bovine embryo development in vitro

Paraoxonase‐1 (PON1) is an enzyme found in serum and follicular fluid that protects cell membrane and circulating lipids against oxidative damage. The aims of this study were to measure the direct effects of recombinant PON1 (rPON1) on bovine oocyte maturation at the molecular level (gene expression) and to measure the carry‐over effects of PON1 on pre‐implantation embryo development in vitro. COCs were submitted to IVM with the addition of 0.0, 0.02, 0.04 and 0.08 mg ml^?1 of rPON1, corresponding to an average PON1 arylesterase enzyme activity of 2.2 ± 0.4, 15.5 ± 1.5, 30.2 ± 3.0 and 57.9 ± 5.0 U ml^?1, respectively. The results indicated that addition of rPON1 during IVM improved embryo development in a dose‐dependent manner as D7 embryo development was 22.2%, 29.4%, 32.2% and 37.0% for the treatment groups, respectively (p = 0.02). In conclusion, addition of PON1 enzyme during IVM exerted dose‐related positive effects on embryo development rates to blastocysts.     

Relationship of Progesterone,Bovine Pregnancy-Associated Glycoprotein-1 and Nitric Oxide with Late Embryonic and Early Fetal Mortalities in Dairy Cows

The aim of the present study was to determine the relationship of progesterone (P4), bovine pregnancy-associated glycoprotein-1 (bPAG-1) and nitric oxide (NO) levels with late embryonic (LEM; day 28 to day 42) and early fetal mortalities (EFM; > day 42 to day 56) in dairy cows. Transrectal ultrasonography (6–8 MHz) was performed in 100 Holstein-Friesian cows at days 28, 42 and 56 after artificial insemination (AI; day 0) to diagnose pregnancy and to monitor the fate of the embryo. After ultrasound scanning of each cow, a milk sample was collected for assessment of P4 by an ELISA test and a blood sample was collected for assessment of bPAG-1, by using a double-antibody radioimmunoassay, and serum NO metabolites (nitrate + nitrite). Based on ultrasonographic examinations and bPAG-1-RIA, 41 of 100 inseminated cows were confirmed pregnant at day 28 after AI. Nine cows suffered of LEM, and 6 cows suffered of EFM and the overall pregnancy loss rate was 36.6% (15/41) between days 28 and 56 of pregnancy. By logistic regression analysis, there were no significant relationships between the level of P4 and bPAG-1 at day 28 after AI and the occurrence of LEM and EFM. Also, there were no significant relationships between the levels of P4 and bPAG-1 at day 42 and the occurrence of EFM. On the other hand, a significant relationship (P<0.05) was found between NO level at day 28 and the occurrence of LEM. In conclusion, measurement of the serum NO concentration at day 28 of pregnancy might help to predict the outcome of pregnancy by day 42 in dairy cows but further studies are needed to confirm this.     

Genetic variability and heterotic groups of Brazilian popcorn populations

The development of successful advanced lines and cultivars in maize is dependent on parental selection and assignment to defined heterotic groups. So, the objectives of this study were to evaluate genetic variability and identify heterotic groups among Brazilian popcorn varieties. Thus, diallel crosses of advanced generations of the popcorn hybrids, IAC 112 and Zélia, and of three open-pollinated popcorn varieties, RS 20, Branco, and SAM were performed. Ten hybrid combinations, the five parents, and five check treatments were arranged in a block design with four replicates in two tropical-zone locations (CWb climate). Both additive and non-additive effects were important for grain yield, plant height, ear height, and husk cover. For popping expansion, only the additive effects were important. Hybrid combinations between the local variety (Branco) and F₂ populations (IAC 112 and Zelia) resulted in the third and fourth highest values for popping expansion. The best grain yields were obtained with hybrid combinations involving SAM. Cultivars Zélia, IAC 112, and RS 20 increased popping expansion, whereas cultivar Branco increased grain yield of hybrid combinations. The following conclusions may be drawn: Brazilian popcorn populations have reduced heterosis and genetic variability to popping expansion in relation commercial cultivars; there is genetic variability among Brazilian popcorn populations that allows the exploitation of additive and non-additive effects for grain yield; it is possible to increase grain yield by using local varieties; but it would be difficult to obtain commercial hybrids from local varieties because they have poor performance for popping expansion.     

Germination and seedling growth of <Emphasis Type="Italic">Dimorphandra jorgei</Emphasis> MF Silva and <Emphasis Type="Italic">Swartzia macrostachya</Emphasis> Benth. var. <Emphasis Type="Italic">riedelii</Emphasis> Cowan

Dimorphandra jorgei MF Silva (Caesalpinaceae) and Swartzia macrostachya Benth. var. riedelii Cowan (Fabaceae) are tree species from the southern Bahian Atlantic Forest, identified for the restoration of degraded areas. The objective of this research is to study their seed germination and seedling growth to develop simple methods for seedling production in small scale nurseries. Because the species have distinct dormancy and germination characteristics, the seeds were submitted to different treatments. D. jorgei seeds were immersed in hot water, scarified and stored for 12 weeks in the laboratory environment (25°C). S. macrostachya seeds were submitted to different combinations of drying and storage in the laboratory environment or refrigerator (4°C). Seedlings were submitted to two light treatments for 3 months: partial shade (52% of full sun) and full shade (12% of full sun). In D. jorgei, the highest germination occurred for scarified seeds (87%) and for seeds stored for 12 weeks and then scarified (69%). In S. macrostachya the undried control seeds had the highest germination, followed by that of the refrigeration storage treatment, without predrying (99% and 97%, respectively). The average values for the growth parameters were greater in full shade for D. jorgei, while S. macrostachya seedlings showed greater growth rates in partial shade. Two months growth is recommended before D. jorgei seedlings are of the sufficient size and leaf number for field planting. On the other hand, a period of 4 months is recommended for S. macrostachya, due to its relatively slow growth.