Identification of reference genes for studies of quantitative gene expression in male and female quail tissues.

In industrial poultry, quail production has gained increasing prominence over the years. It is known that the intensification of genetic studies has contributed greatly to this growth, through techniques, such as analysis of gene expression by PCR, for example. This study aimed to evaluate stability and recommend reference genes for quantitative real-time PCR in different tissues from male and female broiler quails. The stability of 10 housekeeping genes (GAPDH, RPL5, MRPS27, MRPS30, TFRC, HMBS, EEF1, LDHA, B2M, and UBC) by means Bestkeeper, NormFinder, GeNorm softwares with ΔCq method. The tissues analyzed were: heart, thigh muscle, brain, and spleen, considering that they are tissues commonly used in nutrigenomic, immunological, and poultry performance research. As expected, the reference genes tested showed varying stability depending on the tissue evaluated. According to the present study, the most stable housekeeping genes were MRPS30, TFRC, and HMBS in heart; MRPS30, EEF1, and HMBS in thigh muscle; B2M, GAPDH, and UBC in brain; and EEF1, LDHA, and HMBS in spleen. Therefore, it is recommended to be used as reference genes for gene expression studies of male and female quails.

Effects of additional gonadotropin-releasing hormone and prostaglandin F2α treatment to an estradiol/progesterone-based embryo transfer protocol for recipient lactating dairy cows.

This study was designed to evaluate whether the utilization of a second PGF2α treatment at the end of an estradiol/progesterone (E2/P4)-based protocol with or without GnRH at the beginning of the protocol would improve pregnancy rates of lactating Holstein cows assigned to timed embryo transfer. A total of 501 lactating Holstein cows in 5 farms were enrolled in the experiment. Within farm, cows were blocked by parity and, within block, were assigned randomly to (1) insertion of an intravaginal P4 device (controlled internal drug-releasing device; CIDR) and estradiol benzoate on d -11, PGF2α on d -4, CIDR withdrawal and an injection of estradiol cypionate on d -2, and timed embryo transfer on d 7 (1-PGF; n = 164); (2) the same treatments as 1-PGF, but with PGF2α administered on d -4 and -2 (2-PGF; n = 171); and (3) 2-PGF with the addition of a GnRH treatment on d -11 (GnRH+2-PGF; n = 166). Ovaries were scanned by transrectal ultrasonography on d -11, -4, and 7, and blood samples were collected on d -11, -4, 0, and 7 for P4 determination. Treatment comparisons were performed using contrasts. The proportion of cows with a new corpus luteum on d -4 was greater in GnRH+2-PGF cows. Cows in 1-PGF had a greater P4 concentration on d 0 but lesser P4 on d 7 compared with cows in the other groups. Cows assigned to receive 2-PGF (2-PGF and GnRH+2-PGF) had greater estrus expression, and a greater proportion of cows ovulated to estradiol cypionate. No further contrast effects were observed for follicle diameter, double ovulation rate, pregnancy per embryo transfer (P/ET) on d 32 and 60, or pregnancy loss. As P4 concentration on d -4 increased, P/ET on d 60 tended to increase. Cows with P4 ≥3.66 ng/mL on d -4 had greater P/ET on d 32 and 60 than those with P4 below that threshold. Regardless of treatment, cows with P4 concentration ≥3.66 ng/mL also had a greater pregnancy per synchronized protocol (P/SP) on d 60. Also, a P4 concentration on d -4 (low or high) × follicle diameter (continuous) interaction tendency was observed when evaluating P/ET. Although P/ET did not differ among cows with different follicles sizes with reduced P4 concentration on d -4 (<3.66 ng/mL), it increased in cows with larger follicles exposed to increased P4 concentration (≥3.66 ng/mL). When P4 on d 0 was evaluated, P/ET on d 32 and 60 was greater for cows with low (≤0.09 ng/mL) versus high (>0.21 ng/mL) P4; as P4 concentration on d 0 increased, P/ET linearly decreased. In summary, cows with increased P4 concentrations during growth of the ovulatory follicular wave had improved P/ET. Administering a second PGF2α dose reduced P4 concentration on d 0 and increased ovulatory response to the protocol, but no benefits were observed on P/ET or P/SP.

Revisiting the metallothionein genes polymorphisms and the risk of oral squamous cell carcinoma in a Brazilian population.

BACKGROUND: Metallothioneins (MTs) gene polymorphisms have been associated with the ability of free radical scavenging and detoxification of heavy metals leading to cancer development. Our aim was to revisit, in a Brazilian population, single-nucleotide polymorphisms (SNPs) of the MT gene family previously associated with oral squamous cell carcinoma (OSCC). MATERIAL AND METHODS: A case-control investigation with 28 OSCC patients and 45 controls was conducted, using conventional risk factors (tobacco use and alcohol consumption) as covariates. SNPs genotyping for rs8052334 (MT1B), rs964372 (MT1B), and rs1610216 (MT2A) was performed by PCR-RFLP, and SNPs for rs11076161 (MT1A) were analyzed by TaqMan assay. RESULTS: The only SNP associated with increased risk for OSCC was the MT-1A AA genotype (OR = 4.7; p = 0.01). We have also evidenced for the first time a significant linkage disequilibrium between the SNPs of MT-2A and MT-1A in this population with the highest frequency (30%) of the unfavorable haplotype G/A/C/T (rs1610216 / rs11076161 / rs964372 / rs8052334) of MT gene polymorphisms (OR = 6.2; p = 0.04). Interestingly, after removing the effects of conventional risk factors, we have uncovered the significance of the AA genotype of the rs11076161 with increased odds of 19-fold higher towards OSCC development. CONCLUSIONS: This is the first demonstration that a significant linkage disequilibrium among gene polymorphisms of the MT family may affect susceptibility to oral cancer, which is conditioned by the G/A/C/T haplotype (rs1610216/rs11076161/rs964372/ rs8052334) and the MT-1A gene polymorphism has a potential clinical utility for the OSCC risk assessment.

Usefulness of autocidal gravid ovitraps for the surveillance and control of Aedes (Stegomyia) aegypti (Diptera: Culicidae) in eastern Colombia.

In the past decade, new strategies have been developed to control the Aedes aegypti (Diptera: Culicidae) mosquito vector, as well as a broad range of arboviral agents. Vector control surveillance programmes in Puerto Rico and Australia have implemented the Centers for Disease Control and Prevention autocidal gravid ovitrap (AGO), which has had an impact on vector density and, consequently, the epidemiology of arboviral infections. Colombia intends to establish the AGO as a new tool for the surveillance and control of the A. aegypti vector. AGOs were evaluated in a hyperendemic area for dengue virus during an 8-week period in Villavicencio city, eastern Colombia. The results indicated that the AGOs detect a high density of A. aegypti, with positive results for these traps of over 80% and an average catch of six individuals per trap per week. Acceptance of AGOs in the community exceeded 95%, and adherence was around 89%. This study's results demonstrate, for the first time in Colombia, that traps are a useful tool for the surveillance of A. aegypti. Future studies must consider the implementation of AGOs in the region.

Cowden syndrome is a risk factor for multiple neoplasm: a case report.

BACKGROUND: Cowden's syndrome is an autosomal dominant disease with variable penetrance, involving the tumor suppressor phosphatase and tension homolog gene, located on chromosome 10q22-23, responsible for cell proliferation, migration, and cellular apoptosis. Its clinical presentation encompasses mucocutaneous lesions, which are present around 99% of the time; macrocephaly; and cognitive impairment, and it precedes the appearance of neoplasms such as thyroid carcinoma, breast cancer, among others. In addition to these malformations, arteriovenous malformations of the brain and spine, endocrine abnormalities, skeletal defects, and cardiopulmonary lesions may also be found. The relevance of the case is due to the fact that, through a certain phenotype, the patient's genotype can be inferred and thus followed up closely. CASE REPRESENTATION: The clinical case concerns a 28-year-old Caucasian and Portuguese woman with palmar pits, macrocephaly, and cognitive impairment. She was diagnosed with papillary thyroid carcinoma at 22 years of age and proposed total thyroidectomy. At age 27, a pregnancy was diagnosed with a Breast Imaging-Reporting and Data System 2-rated breast lump. After the histological verification, it was concluded that it was a high metastatic breast sarcoma, opting for palliative mastectomy. A genetic evaluation confirmed alteration in the phosphatase and tension homolog gene, confirming Cowden's syndrome. The patient died at age 29 due to neoplastic pathology. CONCLUSION: This report aims to alert to the clinical signs of this entity and the clinical supervision and follow-up of these patients. In order to prevent premature deaths and to improve patient's quality of life, genetic diseases with cancer impact should be diagnosed as early as possible.

Influence of parasite load on the diagnosis and occurrence of eosinophilia in alcoholic patients infected with Strongyloides stercoralis.

Alcoholic patients are more susceptible to Strongyloides stercoralis infection. The chronic use of alcohol raises the levels of endogenous corticosteroids, which regulates the development of larvae and stimulates the differentiation of rhabditiform into infective filariform larvae, thus inducing internal autoinfection. Therefore, early diagnosis is important to prevent severe strongyloidiasis. The aim of this study was to evaluate the efficacy of parasitological methods, according to the parasite load and the number of stool samples, for diagnosis of S. stercoralis infection, as well the peripheral blood eosinophil count in alcoholic patients. A total of 330 patients were included in this study. The diagnosis was established using three parasitological methods: agar plate culture, Baermann-Moraes method and spontaneous sedimentation. Peripheral eosinophilia was considered when the level was >600 eosinophils/mm3. The agar plate culture (APC) had the highest sensitivity (97.3%). However, the analysis of multiple samples increased the sensitivity of all parasitological methods. The sensitivities of the methods were influenced by the parasite load. When the larval number was above 10, the sensitivity of APC was 100%, while in spontaneous sedimentation the sensitivity reached 100% when the larval number was above 50. In the present study, 15.4% of alcoholic patients infected with S. stercoralis (12/78) had increased peripheral blood eosinophil count (above 600 eosinophils/mm3). For an efficient parasitological diagnosis of S. stercoralis infection in alcoholic patients, repeated examination by two parasitological methods must be recommended, including agar plate culture due to its higher sensitivity. Moreover, S. stercoralis infection was associated with eosinophilia, mostly in patients excreting up to 10 larvae/g faeces.

Synthesis, characterization and applications of maghemite beads functionalized with rabbit antibodies.

Magnetic nanoparticles (NPs) have attracted great attention owing to their applications in the biomedical field. In the present work, maghemite (γFe2O3) NPs of 6.5 nm were prepared using a sonochemical method and used to prepare magnetic beads through silanization with 3-aminopropyltrimethoxysilane (APTS). Subsequently, amino groups in the resulting APTS-γFe2O3 beads were converted to carboxylic acid (CARB-γFe2O3) through the succinic anhydride reaction, as confirmed by transmission electron microscopy (TEM), Fourier transform infrared spectroscopy and dynamic light scattering (DLS) measurements. The size of these beads was measured as 12 nm and their hydrodynamic diameter as 490 nm, using TEM analysis and DLS, respectively. The CARB-γFe2O3 beads were further functionalized by immobilizing rabbit antibodies on their surfaces; the immobilization was confirmed by flow cytometry and ionic strength. The samples were further characterized by Mössbauer spectroscopy and DC magnetization measurements. Studies on magnetic relaxivities showed that magnetic beads present great potential for application in MR imaging.

Updating national standards for drinking-water: a Philippine experience.

The latest version of the Philippine National Standards for Drinking-Water (PNSDW) was issued in 2007 by the Department of Health (DOH). Due to several issues and concerns, the DOH decided to make an update which is relevant and necessary to meet the needs of the stakeholders. As an output, the water quality parameters are now categorized into mandatory, primary, and secondary. The ten mandatory parameters are core parameters which all water service providers nationwide are obligated to test. These include thermotolerant coliforms or Escherichia coli, arsenic, cadmium, lead, nitrate, color, turbidity, pH, total dissolved solids, and disinfectant residual. The 55 primary parameters are site-specific and can be adopted as enforceable parameters when developing new water sources or when the existing source is at high risk of contamination. The 11 secondary parameters include operational parameters and those that affect the esthetic quality of drinking-water. In addition, the updated PNSDW include new sections: (1) reporting and interpretation of results and corrective actions; (2) emergency drinking-water parameters; (3) proposed Sustainable Development Goal parameters; and (4) standards for other drinking-water sources. The lessons learned and insights gained from the updating of standards are likewise incorporated in this paper.

Implants composed of digoxin and poly(ε-caprolactone): development, characterization, anti-proliferative and anti-angiogenic activities.

Drug delivery systems could be applied to locally treat cervical cancer, thus preventing the drawbacks of conventional therapy. In this study, anti-proliferative and anti-angiogenic effects of digoxin incorporated into poly(ε-caprolactone) implants were evaluated, aiming at the local treatment of cervical cancer. Implants were characterized, and the in vitro release profile of digoxin was demonstrated. Anti-proliferative and anti-angiogenic activities of digoxin were investigated by using chorioallantoic membrane and human cervix carcinoma (HeLa) cells, respectively. The chemical structure of digoxin and the semi-crystalline nature of poly(ε-caprolactone) were preserved after designing implants. The hydrophobicity of drug and polymer as well as the semi-crystalline structure provided a controlled diffusion of digoxin from implants. Digoxin released from implantable devices exhibited anti-proliferative activity against HeLa cells. The anti-angiogenic effect was also shown. Finally, implants composed of digoxin and poly(ε-caprolactone) could be applied as a therapeutic alternative to treat the early stage of cervical cancer, once they were able to locally control the release of this anti-angiogenic and anti-proliferative drug, minimizing its systemic side effects and toxicity.

Detection of pathogenic Escherichia coli and microbiological quality of chilled shrimp sold in street markets.

UNLABELLED: Foodborne illnesses caused by Escherichia coli are one of the most important gastrointestinal diseases and therefore represent a public health risk. The presence of E. coli in water or in products such as shrimp indicates faecal contamination. However, indicator micro-organisms can be used to evaluate the microbiological quality of food sold in markets. This study focused on detecting isolates of E. coli containing the genes stx1A, stx2A, eae, LTI, STa, STb, aggR and pCVD432 in chilled shrimp sold in street markets in the municipality of São Paulo, Brazil, and to assess the microbiological quality of this product. Enteropathogenic and enterotoxigenic E. coli pathotypes were detected on the surface of two chilled shrimp samples. Salmonella spp. was not isolated. In addition, contamination of surface and muscle of the shrimp samples was found to be correlated. The detection of EPEC and ETEC pathotypes in chilled shrimp sold in street markets in Brazil provides useful epidemiological information for public health authorities to improve food safety and public health. SIGNIFICANCE AND IMPACT OF THE STUDY: Shrimps are crustaceans commonly produced and consumed in Brazil. Specimens of Farfantepenaeus brasiliensis and Litopenaeus schmitti sold in street markets were examined by PCR to detect the presence of Escherichia coli pathotypes (enteropathogenic, enterotoxigenic, enterohemorrhagic and enteroinvasive). EPEC and ETEC strains were detected in whole shrimp. These findings provide useful information for public health authorities to improve the food safety and health of the Brazilian population.

Increased detection of latent tuberculosis by tuberculin skin test and booster phenomenon in early rheumatoid arthritis patients.

The incidence of tuberculosis (TB) in rheumatoid arthritis (RA) patients is up to four times higher when compared to the general population, but their risk increases with the use of TNF-a drugs. Appropriate screening of latent tuberculosis infection (LTBI) and proper management of such cases substantially reduce the incidence of active TB. Tuberculin skin test (TST) is a widely used method for the detection of LTBI. The time of diagnosis of RA as well as the age of the patient might modify the TST performance. We did an observational, comparative study of RA patients with early and established disease, with the objective to know the prevalence of LTBI using the TST and booster test; an induration ≥5 mm was considered reactive. We evaluated 143 patients (83 [58 %] early RA patients). We found 31.3 and 21.7 % TST positivity in early and established RA patients, respectively. With the use of booster test, the positivity increased to 46.5 and 28.8 %, respectively (p = 0.048, OR 1.33, 95 % CI 1.01-1.75). In conclusion, we found that TST and booster test increased LTBI detection in early RA patients, which may suggest that time of RA diagnosis might affect cellular immunity and therefore the TST response.

Effect of adding a gonadotropin-releasing-hormone treatment at the beginning and a second prostaglandin F2α treatment at the end of an estradiol-based protocol for timed artificial insemination in lactating dairy cows during cool or hot seasons of the year.

Our hypothesis was that fertility could be increased in a timed artificial insemination (TAI) protocol based on estradiol (E2) and progesterone (P4) by combining GnRH with E2-benzoate at the start of the protocol to increase circulating P4 during preovulatory follicle development and by using 2 prostaglandin F2α (PGF) treatments at the end to decrease P4 near TAI. Lactating Holstein cows (n=1,808) were randomly assigned during the cool or hot season of the year to receive TAI (d 0) following 1 of 3 treatments: (1) control: controlled internal drug-release insert + 2mg of E2-benzoate on d -11, PGF on d -4, controlled internal drug-release insert withdrawal + 1.0mg of E2-cypionate on d -2, and TAI on d 0; (2) 2PGF: identical to control protocol with addition of a second PGF treatment on d -2; (3) GnRH: identical to 2PGF protocol with addition of a 100-µg GnRH treatment on d -11. Pregnancy diagnoses were performed on d 32 and 60 after TAI. Season had major effects on many reproductive measures, with cool season greater than hot season in percentage of cows with corpus luteum (CL) at PGF (62.9 vs. 56.2%), ovulatory follicle diameter (15.7 vs. 14.8mm), expression of estrus (86.7 vs. 79.9%), ovulation following the protocol (89.7 vs. 84.3%), and pregnancies per artificial insemination (P/AI; 45.4 vs. 21.4%). The GnRH protocol increased percentage of cows with CL (control=56.9%; 2PGF=55.8%; GnRH=70.5%) and P4 at PGF (control=3.28±0.22; 2PGF=3.35±0.22; GnRH=3.70±0.21ng/mL), compared with control and 2PGF protocols. The GnRH protocol increased P/AI at the pregnancy diagnosis at 32d [37.3% (219/595)] and 60d [31% (179/595)] after TAI, compared with control [30.0% (177/604); 25.1% (145/604)], with intermediate results with 2PGF protocol [33.2% (196/609); 28.0% (164/609)]. The positive effects of GnRH treatment on P/AI were only detected during the cool season (GnRH=50.9%; 2PGF=44.2%; control=41.0%) and not during the hot season. In addition, the effect of GnRH was only observed in cows with low P4 (<3ng/mL) at the start of the protocol and not in cows that began the protocol with high P4. Furthermore, presence of CL at PGF interacted with follicle diameter such that cows with a CL at PGF had greater P/AI if they ovulated larger rather than smaller follicles near TAI. Thus, fertility to TAI can be improved by inducing ovulation at the beginning of an E2/P4-based protocol using GnRH treatment, particularly during the cool season of the year and in cows with low P4 at the start of the protocol.

Cytogenetic characterization of species of the family Heptapteridae (Teleostei: Siluriformes) from the Cuiabá and Ivaí River Basins, Brazil.

We cytogenetically characterized three species of Heptapteridae (Pimelodella sp, Pimelodella taenioptera, and Imparfinis schubarti) by investigating the distribution of constitutive heterochromatin and nucleolar organizer regions by silver nitrate impregnation (Ag-NOR) and fluorescence in situ hybridization. Pimelodella sp showed had a diploid number (2n) = 46 chromosomes, 26m + 10sm + 10st, and FN = 92; P. taenioptera, 2n = 52 chromosomes, 26m + 22sm + 4st, and FN = 104; and I. schubarti, 2n = 58 chromosomes, 28m + 28sm + 2st, and FN = 116. The two Pimelodella species had Ag-NORs sites on the submetacentric pair 14, located on the short arm in terminal position. In I. schubarti, the Ag-NORs sites were in an interstitial position on the long arm of the metacentric pair 1. C-banding revealed that Pimelodella sp contained a small amount of constitutive heterochroma-tin, whereas P. taenioptera contained a higher number of heterochromatic regions, in the pericentromeric, interstitial, and telomeric positions. I. schubarti had markers in centromeric and telomeric regions of a few chromosomes, and a large pericentromeric block on pair 1. Fluorochrome chromomycin A3 (CMA3) staining revealed positive signals on pair 14 in both Pimelodella species. Treatment with 4ꞌ,6-diamidino- 2-phenylindole (DAPI) revealed no markings in P. taenioptera, but an interstitial marking on the long arm of pair 14 in Pimelodella sp. In I. schubarti, positive signals of CMA3 were detected in the first pair, but negative signals were detected for DAPI staining. These results con-tribute to the karyotypic description of the less-studied species in the Brazilian Midwest.

Isolation and characterization of yeasts capable of efficient utilization of hemicellulosic hydrolyzate as the carbon source.

Few yeasts have shown the potential to efficiently utilize hemicellulosic hydrolyzate as the carbon source. In this study, microorganisms isolated from the Manaus region in Amazonas, Brazil, were characterized based on their utilization of the pentoses, xylose, and arabinose. The yeasts that showed a potential to assimilate these sugars were selected for the better utilization of lignocellulosic biomass. Two hundred and thirty seven colonies of unicellular microorganisms grown on hemicellulosic hydrolyzate, xylose, arabinose, and yeast nitrogen base selective medium were analyzed. Of these, 231 colonies were subjected to sugar assimilation tests. One hundred and twenty five of these were shown to utilize hydrolyzed hemicellulose, xylose, or arabinose as the carbon source for growth. The colonies that showed the best growth (N = 57) were selected, and their internal transcribed spacer-5.8S rDNA was sequenced. The sequenced strains formed four distinct groups in the phylogenetic tree, and showed a high percentage of similarity with Meyerozyma caribbica, Meyerozyma guilliermondii, Trichosporon mycotoxinivorans, Trichosporon loubieri, Pichia kudriavzevii, Candida lignohabitans, and Candida ethanolica. The discovery of these xylose-fermenting yeasts could attract widespread interest, as these can be used in the cost-effective production of liquid fuel from lignocellulosic materials.

Anti-PR3 and anti-MPO antibodies are not present in sera of patients with pulmonary tuberculosis.

Anti-neutrophil cytoplasmic antibodies (ANCA) are autoantibodies directed to intracellular components of neutrophils and are present in several vasculitic syndromes. Recently, these autoantibodies have been described in other autoimmune disorders as well as in infectious diseases such as tuberculosis (TB). As there are some clinical similarities between TB and granulomatosis with polyangiitis, we searched for ANCA in a group of patients with proven TB. Patients with TB confirmed by chest X-ray and sputum bacilloscopy either before or within 30 days after beginning treatment were included in this study. Anti-MPO and anti-PR3 antibodies were studied using well-standardized ELISA kits (INOVA Diagnostics, Inc.). ANCA were also investigated by indirect immunofluorescence (IIF). Fifty TB patients (26 females, mean age 47.34 ± 17 years) were enrolled in the present study. No patient tested positive for ANCA by IIF, or anti-MPO or anti-PR3 antibodies by ELISA. Although previous studies have shown the presence of ANCA in some infectious diseases, the findings of the present study demonstrated the absence of such antibodies in TB. The discrepancy in the prevalence of ANCA in TB among different studies may be attributed to methodological factors and/or the genetic background of the studied populations.

Nickel, manganese and copper removal by a mixed consortium of sulfate reducing bacteria at a high COD/sulfate ratio.

The use of sulfate-reducing bacteria (SRB) in passive treatments of acidic effluents containing heavy metals has become an attractive alternative biotechnology. Treatment efficiency may be linked with the effluent conditions (pH and metal concentration) and also to the amount and nature of the organic substrate. Variations on organic substrate and sulfate ratios clearly interfere with the biological removal of this ion by mixed cultures of SRB. This study aimed to cultivate a mixed culture of SRB using different lactate concentrations at pH 7.0 in the presence of Ni, Mn and Cu. The highest sulfate removal efficiency obtained was 98 %, at a COD/sulfate ratio of 2.0. The organic acid analyses indicated an acetate accumulation as a consequence of lactate degradation. Different concentrations of metals were added to the system at neutral pH conditions. Cell proliferation and sulfate consumption in the presence of nickel (4, 20 and 50 mg l(-1)), manganese (1.5, 10 and 25 mg l(-1)) and copper (1.5, 10 and 25 mg l(-1)) were measured. The presence of metals interfered in the sulfate biological removal however the concentration of sulfide produced was high enough to remove over 90 % of the metals in the environment. The molecular characterization of the bacterial consortium based on dsrB gene sequencing indicated the presence of Desulfovibrio desulfuricans, Desulfomonas pigra and Desulfobulbus sp. The results here presented indicate that this SRB culture may be employed for mine effluent bioremediation due to its potential for removing sulfate and metals, simultaneously.

Pollen analysis of honey and pollen stored by Melipona (Melikerria) fasciculata Smith, 1854 (Apidae Meliponini), in an Amazon and Cerrado transition area, Maranhão, Brazil.

The identification of meliponicultural flora is fundamental for the preservation of bees, as well as enabling the development of pollinator management and reforestation programs. In this context, this study aimed to investigate the types of pollen found in the honey and pollen pots of the Melipona fasciculata bee in Arari, State of Maranhão, between August 2022 and July 2023. 40 types of pollen were identified, distributed among 18 botanical families, with Fabaceae standing out (46.15% in pollen pots and 37.86% in honey pots), Myrtaceae (11.53% in pollen and 13.51% in honey) and Rubiaceae (7.69% in pollen and 5.40% in honey). Sapindaceae, in the honey pots, accounted for 5.40%. The other families showed lower percentages, 3.84% in the pollen pots and 2.70% in the honey pots. In terms of similarity, two distinct groups were observed in both the pollen and honey pots. In the pollen pots, group A (May-June ~ 97%) and group B (Feb-Mar ~ 99%) stood out, while in the honey pots, group A (Mar-Apr ~ 98%) and group B (Jun-Jul-Sept ~ 98%) showed the highest percentages. These findings highlight the wide range of resources used by the M. fasciculata species, as well asits preference for Fabaceae and Myrtaceae, due to the diversity and availability of trophic resources. An understanding of the meliponicultural flora is essential to support effective conservation strategies, which aim not only to guarantee the survival of the bees, but also to ensure the continued production of honey, a resource of great importance to local communities.

PAHs in franciscana dolphins from the Southwestern Atlantic Ocean: Concentration and maternal transfer assessments.

Polycyclic aromatic hydrocarbons (PAHs) are organic compounds ubiquitous in the environment and known for their toxic, mutagenic, and carcinogenic effects. These compounds can bioaccumulate in the biota and be transferred through trophic webs. The franciscana dolphin (Pontoporia blainvillei), as top predators, can be an environmental sentinels. Thus, this study aimed to provide data about PAHs concentration in their hepatic tissue collected on the coast of Espírito Santo (Franciscana Management Area, FMA Ia), Rio de Janeiro (FMA IIa), and São Paulo states (FMA IIb), in Southeastern Brazil. PAHs were detected in 86 % of franciscana dolphins (n = 50). The highest ∑PAHsTotal median concentration was reported in FMA Ia followed by FMA IIb and FMA IIa (1055.6; 523.9, and 72.1 ng.g-1 lipid weight, respectively). Phenanthrene was detected in one fetus and two neonates, showing maternal transfer of PAHs in these dolphins. Evaluating PAHs with potential toxic effects is of utmost importance for the conservation of a threatened species.

Effect of feeding according to energy balance on performance, nutrient excretion, and feeding behavior of early lactation dairy cows.

The objectives of this study were to evaluate 2 feeding strategies for early lactation cows on performance and efficiency of nutrient utilization. Fifty-eight Holsteins cows were blocked by parity and production during the pretreatment period and then randomly assigned at 21 d postpartum to a control diet [n=29; 16.2% crude protein, 1.64 Mcal of net energy for lactation (NE(L)), 22% starch, and 19% forage neutral detergent fiber (NDF)] or a diet with caloric density manipulated weekly (precision diet; n=29; 16.2% crude protein; 1.59 to 1.68 NE(L); 18 to 26% starch; and 16 to 22% forage NDF) to promote a calculated positive energy balance of 5 Mcal/day. Diets were fed as total mixed rations and precision cows had their diets adjusted individually once a week, by feeding additional grain supplementation from 0 to 25% of daily dry matter (DM) offered, according to the energy balance of the preceding week. Energy balance was calculated daily and then averaged weekly. The study lasted from wk 3 to 19 postpartum, and nutrient digestibility, rumen fluid composition, urinary output, estimates of microbial protein synthesis, and feeding behavior were evaluated between wk 9 and 13 postpartum. Compared with controls, precision cows had similar DM intake (24.3 kg/d), but NE(L) intake tended to be greater primarily between wk 4 and 8 postpartum. Yields of milk (45.2 vs. 41.9 kg/d), milk components, 3.5% fat-corrected milk (44.0 vs. 40.8 kg/d), and energy-corrected milk (43.4 vs. 40.2) were all greater for precision than control cows, resulting in greater energy-corrected milk production per kilogram of diet DM consumed (1.79 vs. 1.72). Precision cows produced more milk calories per kilogram of metabolic weight (0.227 vs. 0.213 Mcal of NE(L)/kg), although the amount of consumed calories partitioned into milk (82.3%) and measures of energy status did not differ between treatments throughout the study. Glucose concentrations were greater throughout the day in precision cows compared with controls at 6 wk, but not 13 wk postpartum. Apparent digestibility of nutrients, composition of rumen fluid, mean and low rumen pH, and estimated rumen microbial N synthesis remained mostly unaltered by treatments. Although precision cows produced more milk true protein, measures of efficiency of dietary N use were not influenced by treatment. On wk 13 postpartum, precision cows consumed a diet with longer NDF particles, which resulted in a tendency for greater intake of NDF >8mm because of less sorting against the long particles than control cows. Meal pattern differed with treatment, and precision cows consumed feed more sparsely throughout the day, spent more time ruminating lying, and had similar meal duration (mean of 36.3 min/meal) compared with control cows, but smaller meal size (3.33 vs. 3.64 kg/meal). Results from the current study indicate that allocating dietary resources according to the individual needs of cows based on energy balance improves lactation performance compared with feeding a single total mixed ration, despite similar average nutrient intake between treatments. Improvements in performance are likely related to allocation of calories based on the needs of the cow and on shifts of feeding behavior that might favor intake of smaller meals.

Targeted progesterone supplementation improves fertility in lactating dairy cows without a corpus luteum at the initiation of the timed artificial insemination protocol.

The objectives of this study were to determine the effect of supplemental progesterone on fertility in lactating dairy cows lacking a corpus luteum (CL) at the initiation of the timed artificial insemination (AI) program. Holstein cows were subjected to the 5-d timed AI program (d -8 GnRH, d -3 and -2 PGF2α, d 0 GnRH and AI). Cows had their ovaries scanned by ultrasonography on d -8 and those bearing a CL were considered to be in diestrus (DI; n=946). Cows that lacked a CL on d -8 were assigned to remain as untreated control (CON; n=234) or receive 2 controlled internal drug release (CIDR) inserts containing progesterone (2CIDR; n=218) from d -8 to -3, as a single insert has been proven insufficient to modulate fertility in cows without CL. Blood was analyzed for progesterone and estradiol concentrations. Pregnancy was diagnosed on d 34 and 62 after AI. Progesterone concentrations during the timed AI program were lowest for CON, intermediate for 2CIDR, and highest for DI. Supplementation increased progesterone concentrations between d -7 and -3 compared with CON (2.65 vs. 0.51 ng/mL). Ovulation to the first GnRH was not affected by treatment. However, a greater proportion of CON and 2CIDR cows had a new CL on d -3 compared with DI cows (66.7 vs. 61.9 vs. 52.0%). In cows with a new CL, the diameter of the ovulatory follicle was larger for CON than 2CIDR, and intermediate for DI (18.7 vs. 16.5 vs. 17.7 mm). Concentrations of estradiol on d -3 did not differ among treatments; however, DI cows had greater estradiol concentrations at AI compared with CON or 2CIDR cows. Pregnancy per AI was less for CON compared with 2CIDR or DI on d 32 (30.8 vs. 46.8 vs. 49.9%) and 64 (28.6 vs. 43.7 vs. 47.3%), indicating that supplementation with progesterone reestablished fertility in cows lacking a CL on d -8. A greater proportion of nonpregnant CON cows had a short reinsemination interval compared with 2CIDR or DI (11.1 vs. 3.5 vs. 5.7%). Treatment did not affect pregnancy loss between d 34 and 62 of gestation. A single ultrasound exam was effective in identifying a low-fertility cohort of cows based on the absence of CL at the first GnRH injection of the timed AI protocol. Progesterone supplementation with 2 CIDR inserts increased progesterone in plasma to 2.65 ng/mL and restored fertility in lactating dairy cows lacking a CL at the initiation of the timed AI program similar to that of cows in diestrus.