Acaricides containing zein nanoparticles: A tool for a lower impact control of the cattle tick Rhipicephalus microplus.

Nanoformulations containing zein nanoparticles (ZN) can promote the stability and protection of molecules with acaricidal activity. The present study sought to develop nanoformulations with ZN associated with cypermethrin (CYPE) + chlorpyrifos (CHLO) + a plant compound (citral, menthol or limonene), characterize them, and verify their efficacy against Rhipicephalus microplus ticks. Additionally, we aimed to assess its safety in nontarget nematodes found in soil at a site subjected to contamination by acaricides. The nanoformulations were characterized by dynamic light scattering and nanoparticle tracking analysis. Nanoformulations 1 (ZN+CYPE+CHLO+citral), 2 (ZN+CYPE+CHLO+menthol), and 3 (ZN+CYPE+CHLO+limonene) were measured for diameter, polydispersion, zeta potential, concentration, and encapsulation efficiency. Nanoformulations 1, 2, and 3 were evaluated in a range from 0.004 to 0.466 mg/mL on R. microplus larvae and caused mortality > 80% at concentrations above 0.029 mg/mL. The commercial acaricide Colosso® (CYPE 15 g + CHLO 25 g + citronellal 1 g) was evaluated also from 0.004 to 0.512 mg/mL and resulted in 71.9% larval mortality at 0.064 mg/mL. Formulations 1, 2, and 3 at 0.466 mg/mL showed acaricidal efficacy of 50.2%, 40.5%, and 60.1% on engorged females, respectively, while Colosso® at 0.512 mg/mL obtained only 39.4%. The nanoformulations exhibited long residual period of activity and lower toxicity to nontarget nematodes. ZN was able to protect the active compounds against degradation during the storage period. Thus, ZN can be an alternative for the development of new acaricidal formulations using lower concentrations of active compounds.

Investigating the benefits of targeted selective treatment according to average daily weight gain against gastrointestinal nematodes in Morada Nova lambs.

The objective of this study was to evaluate a method of targeted selective treatment (TST) of Morada Nova lambs, based on the average daily weight gain (DWG). For this, 114 lambs in the rainy and 102 in the dry season were randomized into three treatments: control (CT), routine (RT, treated every 42 days), and targeted selective (TST, treated according to DWG). Packed cell volume (PCV) and eggs per gram of feces (EPG) tests were performed. The anthelmintic resistance of parasites was assessed by the EPG count reduction test (FECRT), RESISTA-Test©, and molecular test. For CT, RT, and TST groups, the following results were obtained: mean EPG values were 4665.1, 3063.5, and 3462.1 in the rainy season and 4475.1, 1341.7, and 2863.4 in the dry season, respectively; mean PCV values were 32.1, 33.4, and 32.3% in the rainy season and 33.9, 36.0, and 35.1% in the dry season; mean DWG readings were 0.087, 0.101, and 0.094 kg in the rainy season and 0.102, 0.113, and 0.112 kg in the dry season; efficacies of levamisole in FECRT were 66.4, 24.1, and 76.4% in the rainy and 90.7, 12.4, and 64.8% in the dry season, respectively; in the RESISTA-Test©, the LC50 values were 0.482, 1.926, and 0.117 µg.mL-1 in the rainy and 0.437, 0.851, and 0.045 µg.mL-1 in the dry season, respectively; the frequencies of the homozygous-resistant genotype were 57.1, 71.4, and 40.0% in the rainy and 47.8, 55.9, and 41.9% in the dry season. In conclusion, TST reduced the development of resistance through refugia maintenance, without productive losses, contributing to the sustainability of sheep breeding.

Ovine ß-globin gene: A new qPCR for rapid haplotype identification and association with susceptibility to Haemonchus contortus infection.

Two ß-globin allelic haplotypes (A and B) were identified in domestic sheep, wherein animals which are homozygous for ßB allele (BB haplotype) have a deletion of pre-adult ßC-globin and consequently are less tolerant to anemia and hypoxia. Since Haemonchus contortus infection, is associated with severe anemia, studies performed from 1960s to 1990s investigated the association between ß-globin haplotype and resistance against this parasite. However, the findings were controversial, pointing out from increased resistance in animals harboring the ßA allele to inexistence of association. Thus, our study aimed to develop a qPCR for ß-globin haplotype identification, and to evaluate the association between ß-globin haplotype and resistance against H. contortus in a group of sheep submitted to artificial infection with this parasite. A total of 286 lambs of Morada Nova breed were experimentally challenged with 4000 H. contortus L3 and monitored for 112 days from weaning. Significantly improved (p < 0.05) phenotypic profiles (lower fecal egg counts, higher packed cell volume and birthweight) were observed for AA haplotype animals, especially when compared to BB animals, while AB animals were similar to BB. This is the first report of a qPCR assay for ovine ß-globin haplotype identification. In view of significant differences of phenotypic profiles between haplotype groups, the developed qPCR may constitute an important tool for sheep producers to improve genetic selection of parasite resistant animals.

Biological parameters for Rhipicephalus microplus in the field and laboratory and estimation of its annual number of generations in a tropical region.

The present study aimed to evaluate biological parameters of Rhipicephalus microplus in the non-parasitic phase in both field and laboratory conditions. It also aimed to assess correlations between duration (in days) of these parameters and climatic variables (humidity, rainfall, and soil temperature) and to estimate the annual number of generations of R. microplus in a tropical region. The non-parasitic phase of R. microplus in field and laboratory conditions was evaluated throughout the course of two years. A pasture was infested with engorged female of R. microplus, and biological parameters, including female pre-oviposition, female oviposition, egg mass incubation, larval pre-hatching phase, larval maturation, and larval longevity, were evaluated concomitantly with the collection of data on climatic conditions. The same parameters were also evaluated in a climatized chamber in the laboratory. The total duration of the non-parasitic phase in the field was longer in the dry season (1st and 4th life-cycle repetitions) than in the rainy season (2nd, 3rd, and 5th repetitions). Tick biological parameters for the non-parasitic phase in the laboratory were similar to those obtained in the field during the rainy season. The evaluated biological parameters were influenced mainly by environmental and ground-level temperatures, which modified egg mass incubation, larval pre-hatching, and larval longevity periods and, consequently, the total duration of the non-parasitic phase of the tick. The annual number of generations for the tick was estimated at five per year, which is alarming because it represents an increase, and so new studies into strategic control are needed.

Innate Immune Responses Associated with Resistance against Haemonchus contortus in Morada Nova Sheep.

The immune response against Haemonchus contortus infections is primarily associated with the Th2 profile. However, the exact mechanisms associated with increased sheep resistance against this parasite remains poorly elucidated. The present study is aimed at evaluating mediators from the innate immune response in lambs of the Morada Nova Brazilian breed with contrasting H. contortus resistance phenotypes. Briefly, 287 lambs were characterized through fecal egg counts (FEC) and packed cell volume (PCV) after two independent experimental parasitic challenges with 4,000 H. contortus L3. 20 extreme resistance phenotypes (10 most resistant and 10 most susceptible) were selected, subjected to a third artificial infection with 4,000 L3, and euthanized 7 days later. Tissue samples were collected from abomasal fundic and pyloric mucosa and abomasal lymph nodes. Blood samples were collected at days 0 and 7 of the third parasitic challenge. RNA was extracted from tissue and blood samples for relative quantification of innate immune-related genes by RT-qPCR. For the abomasal fundic mucosa, increased TNFα and IL1ß expression levels (P < 0.05) were found in the susceptible animals, while resistant animals had IL33 superiorly expressed (P < 0.05). Higher levels (P < 0.05) of TLR2 and CFI were found in the abomasal pyloric mucosa of resistant animals. TNFα was at higher levels (P < 0.05) in the blood of susceptible lambs, at day 0 of the third artificial infection. The exacerbated proinflammatory response observed in susceptible animals, at both local and systemic levels, may be a consequence of high H. contortus parasitism. This hypothesis is corroborated by the higher blood levels of TNFα before the onset of infection, which probably remained elevated from the previous parasitic challenges. On the other hand, resistant lambs had an enhanced response mediated by TLR recognition and complement activation. Nevertheless, this is the first study to directly associate sheep parasitic resistance with IL33, an innate trigger of the Th2-polarized response.

Viability of Haemonchus placei parasitism in experimentally infected young goats.

The present study aimed to evaluate the viability of Haemonchus placei parasitism in experimentally infected goats. For that, 14 75 days old kids male Saanen kids were placed in one of the four experimental groups: GI - infected with 5000 H. placei L3 (n = 4); GII - infected with 5000 H. contortus L3 (n = 4); GIII - infected with 2500 H. contortus L3 + 2500 H. placei L3 (n = 4), and GIV - control, inoculated with distilled water (n = 2). Each kid received, orally, the infective dose in a single inoculum. Based on daily fecal egg counts, the average pre-patent period was determined as 24 days for H. contortus, and 31 days for H. placei. Regarding the Haemonchus spp. recovered at necropsy, the experimental groups GI, GII, and GIII had, respectively, an average of 25.5, 619.5, and 724.75 (120 H. placei, and 604.75 H. contortus) adult specimens, and no immature forms. Under the conditions of this study, the viability of goat infection by H. placei was confirmed, although, with low susceptibility. Nevertheless, the parasitism of this helminth species was more intense when associated with H. contortus. This fact indicates that in common grazing between cattle and young goats, when the latter end up ingesting both Haemonhcus species, especially in a mixed infection, H. placei may also parasitize them.

Morada Nova sheep breed: Resistant or resilient to Haemonchus contortus infection?

Morada Nova is a Brazilian hair sheep breed that is well adapted to the country's mainly tropical climate and has good potential for meat and leather production. This breed is reported to be resistant to Haemonchus contortus infection, a highly desired characteristic due to the large impact of this parasite on sheep farming. Therefore, the present study aimed to characterize 287 recently weaned Morada Nova lambs and 123 ewes in relation to their resistance against H. contortus. The animals were dewormed and 15 days later artificially infected with 4000 H. contortus L3 (D0). They were individually monitored by periodic assessment of fecal egg count (FEC), packed cell volume (PCV), and live weight (LW). On D42, the sheep were again dewormed and submitted to a new parasitic challenge, following the same scheme. The animals of each category (lambs and ewes) were ranked according to individual mean FEC values, and classified as resistant (R, 20%), intermediate (I, 60%), or susceptible (S, 20%) to H. contortus infection. At weaning, high FEC were observed in all three phenotypes (P>0.05). After the artificial infections, there was a significant difference (P<0.05) among the three lamb phenotypes for the mean FEC (RI > S), and LW (R=I > S). The infection levels (FEC) were negatively correlated with PCV (r = -0.66; P<0.001), and LW (r = -0.30; P<0.001). Despite this, the lambs were resilient, since more than 88% of these animals maintained the PCV above 24%, even when heavily infected. The importance of selective parasite control before weaning to reduce the negative impact on slaughter weight was evidenced, taking into account the high positive correlation between LW at weaning and final LW (r=0.73; P<0.001). The ewes, in turn, were strongly resistant to the parasite. Despite highly significant differences (P<0.001) for mean FEC between phenotypes (R0.05). With the phenotypic characterization performed here, it is possible to introduce procedures for parasite control in Morada Nova flocks, facilitating the target-selective treatment approach. The results of this study can also support improvement of meat production by the Morada Nova breed.

Morada Nova sheep breed: Resistant or resilient to Haemonchus contortus infection?

Morada Nova is a Brazilian hair sheep breed that is well adapted to the country's mainly tropical climate and has good potential for meat and leather production. This breed is reported to be resistant to Haemonchus contortus infection, a highly desired characteristic due to the large impact of this parasite on sheep farming. Therefore, the present study aimed to characterize 287 recently weaned Morada Nova lambs and 123 ewes in relation to their resistance against H. contortus. The animals were dewormed and 15 days later artificially infected with 4000 H. contortus L3 (D0). They were individually monitored by periodic assessment of fecal egg count (FEC), packed cell volume (PCV), and live weight (LW). On D42, the sheep were again dewormed and submitted to a new parasitic challenge, following the same scheme. The animals of each category (lambs and ewes) were ranked according to individual mean FEC values, and classified as resistant (R, 20%), intermediate (I, 60%), or susceptible (S, 20%) to H. contortus infection. At weaning, high FEC were observed in all three phenotypes (P > 0.05). After the artificial infections, there was a significant difference (P < 0.05) among the three lamb phenotypes for the mean FEC (R < I < S), PCV (R > I > S), and LW (R = I > S). The infection levels (FEC) were negatively correlated with PCV (r = -0.66; P < 0.001), and LW (r = -0.30; P < 0.001). Despite this, the lambs were resilient, since more than 88% of these animals maintained the PCV above 24%, even when heavily infected. The importance of selective parasite control before weaning to reduce the negative impact on slaughter weight was evidenced, taking into account the high positive correlation between LW at weaning and final LW (r = 0.73; P < 0.001). The ewes, in turn, were strongly resistant to the parasite. Despite highly significant differences (P < 0.001) for mean FEC between phenotypes (R < I < S), 98% of the ewes maintained FEC below 4000 EPG. Their health was not affected, since PCV and LW did not differ between phenotypes, and these parameters were not significantly correlated with FEC (P > 0.05). With the phenotypic characterization performed here, it is possible to introduce procedures for parasite control in Morada Nova flocks, facilitating the target-selective treatment approach. The results of this study can also support improvement of meat production by the Morada Nova breed.