Molecular detection of Hepatozoon canis in dogs from Ibagué, Tolima.

The genus Hepatozoon consists of apicomplexan protozoans that affect mammals, birds, amphibians, and reptiles. In dogs, the Hepatozoon species include H. canis and H. americanum, which are transmitted by the Rhipicephalus sanguineus tick and cause nonspecific signs, such as fever, weight loss, diarrhea, and blood disorders. These protozoans have a worldwide distribution in Europe, Asia, Africa, and North and South America, including Colombia. The aim of this study was to determine the prevalence and risk factors associated with H. canis in the urban and rural areas of Ibagué, Colombia. Blood samples were collected from 308 dogs (180 rural areas and 128 urban areas). Collected data included dog breed, sex, age, environmental factors, and the presence of ectoparasites. A fragment of the 18S rRNA gene was amplified by PCR for detection of the pathogen and confirmed by sequencing. Among the 308 samples, 14 were positive (14/308, 4.5%) for the presence of H. canis. The partial sequence of the 18S rRNA gene showed identity values >98% with H. canis, forming a cluster with sequences from Latin America. An epidemiological survey found two protective factors: most of the time at home (P=0.055) and overnight stay at home (P=0.03). This is the first molecular study of the prevalence and phylogeny analysis of H. canis in Ibagué, Colombia. The findings may help determine risk factors and enhance our understanding of the geographic distribution of H. canis in Colombia.

Leishmania spp. diagnosis and therapeutic management in a cat from urban area in Ibagué (Colombia).

BACKGROUND: Leishmania spp., a protozoan transmitted by sandflies, widely affects humans and dogs in Colombia, nevertheless feline leishmaniasis (FeL) remains understudied. OBJECTIVE: This study reports a case of feline leishmaniasis in Colombia and its therapeutic management. METHODS: Complete blood count, renal and hepatic serum biochemistry, nodular lesion cytology, FeLV/FIV snap test, abdominal ultrasound, and molecular diagnosis of Leishmania spp. 16 s rRNA gene amplification by real-time-PCR (qPCR), ITS-1 and hsp70 gene by endpoint-PCR and Sanger sequencing were performed. RESULTS: The patient was negative for FIV/FeLV and showed leukocytosis, lymphocytosis, thrombocytopenia, neutrophilia, monocytosis, hypergammaglobulinemia, increased gamma-glutamyl-transferase, cortical nephrocalcinosis, diffuse heterogeneous splenic parenchyma, and cholangitis. Nodular lesion cytology, qPCR and Sanger sequencing confirmed the diagnosis of Leishmania spp. The patient was treated with allopurinol and miltefosine. After treatment, clinical signs disappeared. CONCLUSION: Clinical examination, cytology, and molecular tests allowed a rapid and sensitive FeL diagnosis. Allopurinol and miltefosine improved the clinical condition of the cat.

Egg Quality, Yolk Fatty Acid Profiles from Laying Hens Housed in Conventional Cage and Cage-Free Production Systems in the Andean Tropics.

Egg consumers worldwide have increased their concerns about laying hens' welfare and its impact on final egg product quality. This study compared the egg quality parameters under the conventional cage (CC) and cage-free (CF) egg production systems in the tropics. The study was conducted on a commercial farm in Colombia using Hy-Line Brown pullets, reared under the same conditions for the first 15 wks. At 16 wks, the hens were distributed into two housing systems, CC and CF, on the same farm. The hens were fed the same diet for each phase in both systems and feed intake varied slightly. Egg samples were collected every six wks, from 22 to 82 wks of age. A total of 3960 eggs were analyzed at 11 sampling times. Parameters such as albumen height, egg weight, yolk color, eggshell thickness, eggshell strength, and Haugh units were determined using a DET-6000 machine. At 22 and 82 wks, screening for Salmonella spp. status was conducted using environmental and egg samples. Additionally, at 34, 64, and 82 wks, yolk samples were obtained for fatty acid profiles and crude protein (CP) analysis. The data were analyzed in a completely randomized block design with repeated measures (11 times): mean separation by Student's t-test yolk pigmentation, Haugh Units, and albumen height (p < 0.001) were higher in the CF compared with the CC between 38 and 69 wks of age, and eggs at 63 and 82 wks (p < 0.05) were heavier in the CF compared to the CC. Likewise, eggs from the CC had better eggshell strength from 57 to 82 wks. In the egg yolk fatty acid profile at the 34th wk, the pentadecanoic, palmitic, and heptadecanoic acids had higher concentrations in the CF systems than the CC. At the 64th wk, the egg yolk fatty acids-lauric, myristic, and heptadecanoic-had higher concentrations in the CF; likewise, at the 82nd wk, egg yolks from the CC had higher concentrations of lauric, heptadecanoic, and nervonic fatty acids than the CF. The eggs and environmental samples were negative for Salmonella spp. throughout the whole production phase. These results indicated that the production system might impact internal and external egg quality measures, potentially due to various stressors, including environmental factors or behavior restrictions.

Stress-Related Gene Expression in Liver Tissues from Laying Hens Housed in Conventional Cage and Cage-Free Systems in the Tropics.

Global egg production is mainly based on cage systems, which have been associated with negative effects on the welfare of birds. Stress factors in restrictive production systems can lead to changes in gene transcription and protein synthesis, ultimately impacting the quality of poultry products. The liver serves various metabolic functions, such as glycogen storage, and plays a crucial role in animals' adaptation to environmental changes. Consequently, both internal and external conditions can influence liver functions. The aim of this study was to evaluate the gene expression of AGP, CRP, NOX4, SOD1, CAT, GPX1, SREBF1, and FXR in the liver of laying hens under two different production systems. Liver tissues from Hy-Line Brown hens housed in conventional cage and cage-free egg production systems at 60 and 80 weeks of production were used. mRNA transcript levels were determined by qPCR using the relative quantification method and ACTB as the reference gene. AGP, SOD1, and SREBF1 gene expressions were significantly higher in the conventional cage group at the 60 weeks of production. Furthermore, the mRNA levels of transcripts related to oxidative stress and lipid metabolism were higher in the group of laying hens housed in conventional cages compared to those in cage-free systems. These results suggest differential gene expression of genes related to oxidative stress in liver tissues from hens housed in conventional cages compared to cage-free systems. The conditions of the egg production system can impact the gene expression of oxidative stress and lipid synthesis genes, potentially leading to changes in the metabolism and performance of hens, including egg quality.

Virulence genes identification in Salmonella enterica isolates from humans, crocodiles, and poultry farms from two regions in Colombia.

Background and Aim: Salmonella spp. is frequently found in the digestive tract of birds and reptiles and transmitted to humans through food. Salmonellosis is a public health problem because of pathogenicity variability in strains for virulence factors. This study aimed to identify the virulence genes in Salmonella isolates from humans, crocodiles, broiler cloacas, and broiler carcasses from two departments of Colombia. Materials and Methods: This study was conducted on 31 Salmonella enterica strains from humans with gastroenteritis (seven), crocodiles (seven), broiler cloacas (six), and broiler carcasses (12) from Tolima and Santander departments of Colombia, belonging to 21 serotypes. All samples were tested for Salmonella spp. using culture method on selective and non-selective mediums. Extraction of genomic DNA was performed from fresh colonies, DNA quality was verified by spectrophotometry and confirmed by amplification of InvA gene using conventional polymerase chain reaction (PCR). bapA, fimA, icmF, IroB, marT, mgtC, nlpI, oafA, pagN, siiD, spvC, spvR, spvB, Stn, and vexA genes were amplified by PCR. Results: The most prevalent gene was bapA (100%), followed by marT (96.77%), mgtC (93.55%), and fimA (83.87%). Likewise, IroB (70.97%), Stn (67.74%), spvR (61.29%), pagN (54.84%), icmF (54.8%), and SiiD (45.16%) were positive for more than 50% of the strains. Furthermore, none of the isolates tested positive for the vexA gene. Salmonella isolates presented 26 virulence profiles. Conclusion: This study reported 14 virulence genes in Salmonella spp. isolates from humans with gastroenteritis, crocodiles, and broiler cloacas and carcasses. The distribution of virulence genes differed among sources. This study could help in decision-making by health and sanitary authorities.

Microfilariae infection by Acanthocheilonema reconditum and Dirofilaria immitis and their molecular detection in a dog with lymphoma: Case report.

Objective: Microfilariae parasites are common in tropical regions, and some species are reported as potentially zoonotic. The diagnosis of filarial infection in dogs by cytology or hematologic techniques showed lower sensibility and specificity, which may result in misdiagnosis. Thus, molecular techniques seem to be an alternative to identifying and detecting microfilariae infections. On the other hand, lymphoma is one of the main tumors in domestic animals, with a high prevalence in domestic canines. This study aims to report a mixed infection with microfilariae in a dog with lymphoma, emphasizing its diagnosis and the possible role of this infection in the development of the neoplasia. Materials and Methods: An 8-year-old male mixed breed dog was referred to consultation due to the presence of lethargy, recumbency, skin ulceration lesions, nonspecific pain manifestations, emesis, myoclonus in the left temporalis muscle, and seizures. Routine blood and biochemistry tests were normal, and cytology of the skin evidenced a microfilariae infection. The dog died due to a cardiorespiratory arrest, and tissue sampling was done for histopathology and molecular analysis at the necropsy examination. Results: Skin lesions were related to a microfilarial pyogranuloma related to Acanthocheilonema reconditum. Histopathology of the spleen and liver revealed a diffuse lymphoma composed of blast cells and large lymphocytes, distributed diffusely in the parenchyma and surrounding the vasculature. In the skin, microfilariae were seen in some superficial capillaries. Conclusion: This study describes a microfilariae mixed infection with A. reconditum and Dirofilaria immitis in a dog with a lymphoma and its molecular detection. To the knowledge of the authors, this is the first report of a mixed microfilariae infection in a tumor of a dog and highlights the use of molecular techniques, i.e., polymerase chain reaction, for an accurate diagnosis.

Molecular Characterization of Neurogranin (NRGN) Gene from Red­Bellied Pacu (Piaractus brachypomus).

Neurogranin (NRGN) is a small brain protein expressed in various telencephalic areas and plays an essential role in synaptic plasticity by regulating the availability of calmodulin (CaM). The study aims to characterize the neurogranin gene in Colombian native fish, red-bellied pacu, Piaractus brachypomus, its basal tissue expression and differential expression in brain injury and sublethal toxicity by organophosphates. NRGN gene contains an open reading frame of 183 nucleotides encoding for 60 amino acids. Bioinformatics analysis showed an IQ motif necessary in the interaction with CaM. NRGN mRNA was detected in tissues with higher expression in brain, gills, and head kidney. In brain regions, NRGN showed high expression in the telencephalon (TE) and olfactory bulb (OB). In the sublethal toxicity experiment, NRGN mRNA was upregulated in individuals under organophosphate exposure in the OB and optic chiasm (OC). In brain injury experiment, NRGN showed upregulation at 14 days in OC and at 24 h and 7 days in TE. These findings demonstrate the differential expression of NRGN under different experimental conditions which make it a candidate for a biomarker in the brain of P. brachypomus.

Molecular Characterization of High Mobility Group Box 1a (HMGB1a) Gene in Red-Bellied Pacu, Piaractus brachypomus.

High mobility group box 1 (HMGB1) is a chromosomal protein in the nucleus and a potent extracellular proinflammatory cytokine, widely described in mammals, nevertheless, with scarce reports in fish. In this study, full open reading frame of HMGB1a gene from Piaractus brachypomus is reported as well as its molecular characterization, including tissue gene expression. At predicted protein level, HMGB1a showed similarities with its orthologs in teleosts and higher vertebrates. The relative gene expression of HMGB1a mRNA was measured in several tissues including the brain, where a differential expression appeared in brain regions, i.e., higher expression in the cerebellum and telencephalon. In addition, in an assay of sublethal exposure to chlorpyrifos, upregulation of HMGB1a was detected in optic chiasm. Furthermore, in a traumatic brain injury model, upregulation of HMGB1a expression was evident 24 hours after lesion and remained higher up to 14 days. These findings suggest a role for HMGB1a in brain damage and its candidature as biomarker of brain injury; however, more studies are required to elucidate the functions of HMGB1a and its regulation in P. brachypomus.

Identification of breed-specific genomic variants in Colombian Creole pig breeds by whole-genome sequencing.

Dissecting genetic variation of local breeds is important for the success of conservation. In this research, we investigated the genomic variation of Colombian Creole (CR) pigs, with a focus on the breed-specific variants in the exonic region of 34 genes with reported effects on adaptive and economic traits. Seven individuals of each of the three CR breeds (CM, Casco de Mula; SP, San Pedreño; and ZU, Zungo) were whole-genome sequenced along with 7 Iberian (IB) pigs and 7 pigs of each of the four most used cosmopolitan (CP) breeds (Duroc, Landrace × Large White, and Pietrain). Molecular variability in CR (6,451,218 variants; from 3,919,242, in SP, to 4,648,069, in CM) was comparable to that in CP, but higher than in IB. For the investigated genes, SP pigs displayed less exonic variants (178) than ZU (254), CM (263), IB (200), and the individual CP genetic types (201 to 335). Sequence variation in these genes confirmed the resemblance of CR to IB and indicates that CR pigs, particularly ZU and CM, are not exempt from selective introgression of other breeds. A total of 50 exonic variants were identified as being potentially specific to CR, including a high-impact deletion in the intron between exons 15 and 16 of the leptin receptor gene, which was only found in CM and ZU. The identification of breed-specific variants in genes related to adaptive and economical traits can bolster the understanding of the role of gene-environment interactions on local adaptation and points the way for effective breeding and conservation of CR pigs.

Expression profiling of heat shock protein genes in whole blood of Romosinuano cattle breed.

Background and Aim: Heat shock proteins are highly conserved proteins that work as molecular chaperones expressed in response to thermal stress. This study aimed to determine the expression profile of genes related to the heat stress response in whole blood obtained from the Romosinuano creole breed. Materials and Methods: Real-time polymerase chain reaction was performed to analyze the transcript of hsp90, hsp70, hsp60, and hsf1 in the whole blood of Romosinuano under different temperature-humidity indices (THIs). Results: The expression levels of the hsp70 and hsf1 genes at the high-THI level were higher (p = 0.0011 and p = 0.0003, respectively) than those at the low-THI level. In addition, no differences in the expression levels of the hsp60 and hsP90 genes were detected between the two THIs. Conclusion: The overexpression of hsf1 and hsp70 genes play an important role in protecting cells from damage induced by heat stress.

Identification of reference genes for expression studies in the liver and spleen of laying hens housed in cage and cage-free systems.

Background: The liver and spleen play a pivotal role in metabolism and immune response. During stress, neuroendocrine response induces changes in gene expression, and its assessment demands the validation of the stability of the reference genes to perform relative gene expression experiments. Aim: The objective of this study was to determine the expression stability of four reference genes (GAPDH, ACTB, RNA18S, and HMBS) in the liver and spleen tissues from laying hens housed in a conventional cage (CC) and cage-free (CF) egg production systems. Methods: Liver and spleen from Hy-Line Brown hens housed in CC and CF egg production systems were used. mRNA transcript levels were determined by quantitative polymerase chain reaction (qPCR), and the gene expression stability was evaluated using geNorm, BestKeeper, and NormFinder algorithms. Results: The most stable gene from liver tissue was ACTB in CC, CF, and CC-CF groups (overall data). In the spleen, the most stable genes were GAPDH (CC), HMBS (CF), and ACTB (CC-CF). Conclusion: The ACTB gene was the most stable gene in the liver, and GAPDH and HMBS genes were stable in spleen tissues that could be used for the normalization in qPCR experiments performed in liver and spleen tissues of laying hens housed CC and CF production systems.

First study on microscopic and molecular detection of Acanthocheilonema reconditum and Leishmania infantum coinfection in dogs in Southwest Colombia.

Background and Aim: Canine vector-borne diseases represent an important issue for the welfare and health of animals, but also have great zoonotic potential. These diseases are caused by bacteria, nematodes such as filariae, and other parasites such as Leishmania spp. Given the difficulty in differentiating common microfilariae in dogs by microscopy and serological methods, molecular techniques such as polymerase chain reaction (PCR) and sequencing should be valuable for reaching a reliable diagnosis. This study aimed to use microscopy and PCR to identify the microfilarial species in dogs from Valle del Cauca, Colombia, and a possible association with Leishmania infantum parasites. Materials and Methods: This study was conducted on 270 dogs from Pradera and Florida municipalities. Microfilariae were detected in dogs by optical microscopy and amplification with 5.8S-ITS2-28S. Species identification was achieved through the amplification of the gene cytochrome oxidase I (COX1). Results: Microscopic detection of microfilariae was possible in 4.81% (13/270) of the dogs. In addition, by PCR of COX1 and Sanger sequencing of ITS2, Acanthocheilonema reconditum was identified as the circulating microfilarial species in 12 dogs, coinfecting with the species L. infantum (Leishmania donovani complex). Conclusion: To the best of our knowledge, this is the first report on A. reconditum and L. infantum mixed infection in dogs in Colombia, particularly in the Valle del Cauca.

Molecular Detection of Virulence Factors in Salmonella serovars Isolated from Poultry and Human Samples.

Salmonellosis is a common infectious disease in humans caused by Salmonella spp., which in recent years has shown an increase in its incidence, with products of avian origin being a common source of transmission. To present a successful infective cycle, there are molecular mechanisms such as virulence factors that provide characteristics that facilitate survival, colonization, and damage to the host. According to this, the study aims to characterize the virulence factors of Salmonella spp. strains isolated from broilers (n = 39) and humans (n = 10). The presence of 24 virulence genes was evaluated using end-point PCR. All the strains of Salmonella spp. isolated from broiler chickens revealed presence of 7/24 (29, 16%) virulence genes (lpfA, csgA, sitC, sipB, sopB, sopE, and sivH). Regarding the strains isolated from cases of gastroenteritis in humans, all strains contained (14/24, 58, 33%) virulence genes (lpfA, csgA, pagC, msgA, spiA, sitC, iroN, sipB, orgA, hilA, sopB, sifA, avrA, and sivH). In summary, the presence of virulence genes in different strains of Salmonella isolated from broilers and humans could be described as bacteria with potential pathogenicity due to the type and number of virulence genes detected. These findings are beneficial for the pathogenic monitoring of Salmonella in Colombia.

Thermoprotective molecules: Effect of insulin-like growth factor type I (IGF-1) in cattle oocytes exposed to high temperatures.

The adverse effects of heat stress (HS) on the welfare and productivity of cattle are the result of the associated hyperthermia and the physiological and behavioral mechanisms performed by the animal to regulate body temperature. The negative effects of HS on in vitro oocyte maturation and in vitro bovine embryo production have been reported; being one of the major concerns due to economic and productive losses, and several mechanisms have been implemented to reduce its impact. These mechanisms include supplementation of the medium with hormones, adjuvants, identification of protective genes, among others. This review aims to explore the cellular and molecular mechanisms of insulin-like growth factor-1 (IGF-1) during in vitro and in vivo maturation of bovine oocytes and its thermoprotective effect under HS. Although the supplementation of the culture medium during oocyte maturation with IGF-1 has been implemented during the last years, there are still controversial results, however, supplementation with low concentration showed a positive effect on maturation and thermoprotection of oocytes exposed to higher temperatures. Additionally, IGF-1 is involved in multiple cellular pathways, and it may regulate cell apoptosis in cases of HS and protect oocyte competence under in vitro conditions.

Diversity and distribution of macrofungi (Ascomycota and Basidiomycota) in Tolima, a Department of the Colombian Andes: an annotated checklist.

Background: Macrofungi are classified in the phylum Ascomycota and Basidiomycota and they are very important from an ecological and economic point of view. Most studies of fungi in Colombia have been carried out mainly in the Andean Region, especially in the Departments of Antioquia, Valle del Cauca and Cundinamarca. However, other Departments in the Andean Region, like Tolima, located in the Cordillera Central, are well documented for plants (4,797 species) and animals (2,983 species), but very poorly documented in terms of knowledge of fungal diversity. New information: This study provides a compiled and annotated checklist of all known macrofungi in the Department of Tolima, based on published literature and on the identification of new specimens collected from five localities of the Department. All records were updated taxonomically and we include detailed information on the localities in which they are distributed in the Department. The list includes 164 taxa distributed in 15 orders (Agaricales, Polyporales, Russulales, Boletales, Hymenochaetales, Xylariales, Auriculariales, Thelephorales, Cantharellales, Hypocreales, Pezizales, Gloeophyllales, Phallales, Tremellales, Dacrymycetales) and eighteen records in a doubtful taxa section. We present 26 new reports, 19 for Tolima and nine for Colombia. We also provide genetic and phylogenetic evidence of the occurrence of Gloeoporustelephoroides and Podoscyphavenustula in Colombia. This checklist provides the basis for future studies on species diversity and taxonomy in Tolima, by identifying the least studied taxa and ecosystems and conservation priorities.

Molecular Characterization of Salmonella spp. Isolates from Wild Colombian Babilla (Caiman crocodilus fuscus) Isolated In Situ.

Salmonella enterica is a pathogen capable of colonizing various environments, including the intestinal tract of different animals such as mammals, birds, and reptiles, which can act as carriers. S. enterica infection induces different clinical diseases, gastroenteritis being the most common, which in some cases, can evolve to septicemia and meningitis. Reptiles and amphibians have been reported as a reservoir of Salmonella, and transmission of the pathogen to humans has been documented. This study aimed to determine the presence of virulence genes and characterize the genotypic antibiotic resistance profile in Salmonella strains isolated from Caiman crocodilus fuscus obtained in situ (natural habitat) in Prado, Tolima, Colombia in a previous study and stored in a strain bank in our laboratory. Fifteen Salmonella strains were evaluated through endpoint PCR to determine the presence of resistance genes and virulence genes. The genes blaTEM, strB, and sul1 were detected in all the strains that confer resistance to ampicillin, streptomycin, and sulfamethoxazole, as well as the virulence genes invA, pefA, prgH, spaN, tolC, sipB, sitC, pagC, msgA, spiA, sopB, sifA, lpfA, csgA, hilA, orgA, iroN, avrA, and sivH, indicating the possible role of babilla (Caiman crocodilus fuscus) as a carrier of multidrug-resistant bacteria.

Effect of estrous cycle phases on gene expression in bovine oviduct epithelial cells.

Background and Aim: The oviduct environment is of particular importance because it is the site of fertilization and early embryo development. The oviduct, as a component of the reproductive system, responds to ovarian hormone (estradiol [E2] and progesterone [P4]) stimuli depending on the estrous cycle phase. This study aimed to elucidate the effect of estrous cycle phases (follicular and early and late luteal phases) on gene expression patterns in bovine oviduct epithelial cells (BOECs). Materials and Methods: Oviducts were obtained from healthy slaughterhouse animals, corresponding to ipsilateral ovaries with dominant follicles or corpus luteum during early and late luteal phases. BOECs were recovered from the isthmus (IST) and ampulla (AMP), and the expression patterns of genes related to cytokinesis and mitosis mechanisms (rho-associated coiled-coil containing protein kinase and cellular communication network factor 2 [CCN2]), growth factors (insulin-like growth factor-binding protein 3, epidermal growth factor receptor [EGFR], vascular endothelial growth factor A, and EGFR), antioxidant mechanisms (glutathione peroxidase 4 [GPX4]), apoptosis (B-cell lymphoma 2), complement component (C3), energy metabolism (aldose reductase gene family 1-member b1 [AKRIB1] and solute carrier family 2), hormone receptors (estrogen receptor 1 and luteinizing hormone/choriogonadotropin receptor), and specific glycoproteins (oviductal glycoprotein 1) were analyzed. Results: High P4 levels (late luteal phase) affected the expression of important genes related to antioxidant mechanisms (GPX4), energy metabolism (AKRIB1), growth factors (IGBP3 and EGFR), and cell growth regulation (CCN2) in the AMP. Low P4 levels (early luteal phase) affected the expression of AKR1B1, IGBP3, and CCN2. In addition, estrogen likely had an effect on OVPGP expression in the cattle oviduct. Conclusion: Differential gene expression patterns of BOECs in the AMP during the luteal phase (antioxidant mechanisms, energy metabolism, growth factors, and immunological regulators) and in the IST during the follicular phase (glycoproteins) may influence their renewal and population proportions, modulating the oviduct environment as well as gamete and embryo physiology.

First study on response of astrocytes in alevines of red-bellied pacu (Piaractus brachypomus) to subchronic exposure to chlorpyrifos and trichlorfon.

Background and Aim: Organophosphate pesticides (OPs) used in agricultural production pose environmental and public health risks whenever non-target organisms are exposed to them. Oxon-type OPs, such as trichlorfon (TCF) and chlorpyrifos (CPF), are frequently used in Colombia and have been detected in water bodies in the vicinity of croplands; however, their effect on aquatic organisms, especially fish, is largely unknown. The neurotoxicity of OPs includes inhibition of esterase enzymes, neuronal damage, and increased glial reactivity. This study aimed to assess the astrocytic response in the brain tissue of juvenile red-bellied pacu (Piaractus brachypomus) exposed to TCF and CPF. Materials and Methods: A 25-day subchronic assay was conducted in which juvenile red-bellied pacu were exposed to CPF and TCF. After 25 days of exposure, the fish were killed and brain samples were collected and processed for immunohistochemistry to assess the morphology and reactivity of astrocytes; glial acidic fibrillary protein was used as a biomarker. Results: The brain samples from animals under subchronic exposure to OPs for 25 days showed higher cellular density as well as changes in astrocyte phenotype characterized by shortening of cytoplasmic projections, hypertrophy, and ameboid morphology compared to those from nonexposed animals. Similarly, astrocyte hyperreactivity was detected in the optic tectum and medial longitudinal fasciculus of the exposed group. Conclusion: Immunoreactivity of brain glial cells under subchronic exposure to OPs measured through immunohistochemical tests as well as OPs-induced neuropathology may be useful as a biomarker for monitoring environmental pollution. The results also indicate that P. brachypomus is a suitable biomonitoring model for studying neurotoxicological and neurodegenerative diseases.

Survey of porcine circovirus type 2 and parvovirus in swine breeding herds of Colombia.

BACKGROUND: One of the consequences of the presentation of reproductive failures in sows is the economic losses in production because it alters the estimated values of the volume of production, decreasing the productivity of the farm. Porcine circovirosis by porcine circovirus 2 (PCV2) has been associated with reproductive disorders, and porcine parvovirus (PVP) is one of the pathological agents most related to the presentation of reproductive failure in pigs. In Colombia, there are reports of the presence of PCV2 through molecular techniques, and PVP through serum tests; however, in the department of Tolima, the prevalence of these two viruses is unknown. OBJECTIVE: In this study, the aim was to establish a report of the prevalence of viruses in five municipalities of the department of Tolima-Colombia. METHODS: Blood samples from 150 breeding sows of five municipalities in Tolima, Colombia, were obtained. Quantitative polymerase chain reaction (qPCR) was used to detect the PCV2 and PVP virus in the blood samples followed by PCR and sequencing of 16 PCR products of the amplification of the cap gene of PCV2. A phylogenetic tree was constructed to identify the genotype of the PCV2 virus. RESULTS: The presence of PCV2d in sows was detected in 135 samples (90%), as well as the identification of PVP in 2.6% of the samples. In addition, the phylogenetic analysis showed that 16 isolates were the PCV2d2 genotype. CONCLUSION: PCV2d and PVP were found to coinfect the females, and the identification of variability in regions in the predicted amino acid sequence of the PCV2 capsid may be associated with virus pathogenicity.

Molecular characterization of myelin basic protein a (mbpa) gene from red-bellied pacu (Piaractus brachypomus).

BACKGROUND: Myelin basic protein (MBP) is one of the most important structural components of the myelin sheaths in both central and peripheral nervous systems. MBP has several functions including organization of the myelin membranes, reorganization of the cytoskeleton during the myelination process, and interaction with the SH3 domain in signaling pathways. Likewise, MBP has been proposed as a marker of demyelination in traumatic brain injury and chemical exposure. METHODS: The aim of this study was to molecularly characterize the myelin basic protein a (mbpa) gene from the Colombian native fish, red-bellied pacu, Piaractus brachypomus. Bioinformatic tools were used to identify the phylogenetic relationships, physicochemical characteristics, exons, intrinsically disordered regions, and conserved domains of the protein. Gene expression was assessed by qPCR in three models corresponding to sublethal chlorpyrifos exposure, acute brain injury, and anesthesia experiments. RESULTS: mbpa complete open reading frame was identified with 414 nucleotides distributed in 7 exons that encode 137 amino acids. MBPa was recognized as belonging to the myelin basic protein family, closely related with orthologous proteins, and two intrinsically disordered regions were established within the sequence. Gene expression of mbpa was upregulated in the optic chiasm of the chlorpyrifos exposed fish in contrast to the control group. CONCLUSIONS: The physicochemical computed features agree with the biological functions of MBP, and basal gene expression was according to the anatomical distribution in the tissues analyzed. This study is the first molecular characterization of mbpa from the native species Piaractus brachypomus.