Fungal Vaccine Development: State of the Art and Perspectives Using Immunoinformatics.

Fungal infections represent a serious global health problem, causing damage to health and the economy on the scale of millions. Although vaccines are the most effective therapeutic approach used to combat infectious agents, at the moment, no fungal vaccine has been approved for use in humans. However, the scientific community has been working hard to overcome this challenge. In this sense, we aim to describe here an update on the development of fungal vaccines and the progress of methodological and experimental immunotherapies against fungal infections. In addition, advances in immunoinformatic tools are described as an important aid by which to overcome the difficulty of achieving success in fungal vaccine development. In silico approaches are great options for the most important and difficult questions regarding the attainment of an efficient fungal vaccine. Here, we suggest how bioinformatic tools could contribute, considering the main challenges, to an effective fungal vaccine.

Paracoccidioides lutzii Formamidase Contributes to Fungal Survival in Macrophages.

Nitrogen is a crucial nutrient for microorganisms that compose essential biomolecules. However, hosts limit this nutrient as a strategy to counter infections, therefore, pathogens use adaptive mechanisms to uptake nitrogen from alternative sources. In fungi, nitrogen catabolite repression (NCR) activates transcription factors to acquire nitrogen from alternative sources when preferential sources are absent. Formamidase has been related to nitrogen depletion in Aspergillus nidulans through formamide degradation to use the released ammonia as a nitrogen source. In Paracoccidioides spp., formamidase is highly expressed in transcriptomic and proteomic analyses. Here, we aim to investigate the importance of formamidase to Paracoccidioides lutzii. Thereby, we developed a P. lutzii silenced strain of fmd gene (AsFmd) by antisense RNA technology using Agrobacterium tumefaciens-mediated transformation (ATMT). The AsFmd strain led to increased urease expression, an enzyme related to nitrogen assimilation in other fungi, suggesting that P. lutzii might explore urease as an alternative route for ammonia metabolism as a nitrogen source. Moreover, formamidase was important for fungal survival inside macrophages, as fungal recovery after macrophage infection was lower in AsFmd compared to wild-type (WT) strain. Our findings suggest potential alternatives of nitrogen acquisition regulation in P. lutzii, evidencing formamidase influence in fungal virulence.

ß-Mercaptoethanol in culture medium improves cryotolerance of in vitro-produced bovine embryos.

The objective of this study was to investigate the effects of adding ß-mercaptoethanol (ßME) to culture medium of bovine in vitro-produced (IVP) embryos prior to or after vitrification on embryo development and cryotolerance. In Experiment I, Day-7 IVP blastocysts were vitrified and, after warming, cultured in medium containing 0, 50 or 100 µM ßME for 72 h. Embryos cultured in 100 µM ßME attained higher hatching rates (66.7%) than those culture in 0 (47.7%) and 50 (52.4%) µM ßME. In Experiment II, IVP embryos were in vitro-cultured (IVC) to the blastocyst stage in 0 (control) or 100 µM ßME, followed by vitrification. After warming, embryos were cultured for 72 h (post-warming culture, PWC) in 0 (control) or 100 µM ßME, in a 2 × 2 factorial design: (i) CTRL-CTRL, control IVC and control PWC; (ii) CTRL-ßME, control IVC and ßME-supplemented PWC; (iii) ßME-CTRL, ßME-supplemented IVC and control PWC; or (iv) ßME-ßME, ßME-supplemented IVC and ßME-supplemented PWC. ßME during IVC reduced embryo development (28.0% vs. 43.8%) but, following vitrification, higher re-expansion rates were seen in ßME-CTRL (84.0%) and ßME-ßME (87.5%) than in CTRL-CTRL (71.0%) and CTRL-ßME (73.1%). Hatching rates were higher in CTRL-ßME (58.1%) and ßME-ßME (63.8%) than in CTRL-CTRL (36.6%) and ßME-CTRL (42.0%). Total cell number in hatched blastocysts was higher in ßME-ßME (181.2 ± 7.4 cells) than CTRL-CTRL (139.0 ± 9.9 cells). Adding ßME to the IVC medium reduced development but increased cryotolerance, whereas adding ßME to the PWC medium improved embryo survival, hatching rates, and total cell numbers.

Challenges in Serologic Diagnostics of Neglected Human Systemic Mycoses: An Overview on Characterization of New Targets.

Systemic mycoses have been viewed as neglected diseases and they are responsible for deaths and disabilities around the world. Rapid, low-cost, simple, highly-specific and sensitive diagnostic tests are critical components of patient care, disease control and active surveillance. However, the diagnosis of fungal infections represents a great challenge because of the decline in the expertise needed for identifying fungi, and a reduced number of instruments and assays specific to fungal identification. Unfortunately, time of diagnosis is one of the most important risk factors for mortality rates from many of the systemic mycoses. In addition, phenotypic and biochemical identification methods are often time-consuming, which has created an increasing demand for new methods of fungal identification. In this review, we discuss the current context of the diagnosis of the main systemic mycoses and propose alternative approaches for the identification of new targets for fungal pathogens, which can help in the development of new diagnostic tests.

An update on the occurrence of Paracoccidioides species in the Midwest region, Brazil: Molecular epidemiology, clinical aspects and serological profile of patients from Mato Grosso do Sul State.

BACKGROUND: Paracoccidioidomycosis (PCM) is a systemic and endemic fungal infection in Latin American, mainly in Brazil. The majority of PCM cases occur in large areas in Brazil, comprising the South, Southeast and Midwest regions, with the latter demonstrating a higher incidence of the species Paracoccidioides lutzii. METHODOLOGY AND MAIN FINDINGS: This study presents clinical, molecular and serological data of thirteen new PCM cases during 2016 to 2019 from the state of Mato Grosso do Sul, located in the Midwest region, Brazil. From these thirteen cases, sixteen clinical isolates were obtained and their genomic DNAs were subjected to genotyping by tub1 -PCR-RFLP. Results showed Paracoccidioides brasiliensis sensu stricto (S1) (11/16; 68.8%), Paracoccidioides restrepiensis (PS3) (4/16; 25.0%) and P. lutzii (1/16; 6.2%) as Paracoccidiodes species. Therefore, in order to understand whether the type of phylogenetic species that are circulating in the state influence the reactivity profile of serological tests, we performed double agar gel immunodiffusion (DID), using exoantigens from genotyped strains found in this series of PCM cases. Overall, our DID tests have been false negative in about 30% of confirmed PCM cases. All patients were male, most with current or previous rural activity, with ages ranging from 17 to 59 years, with 11 patients (84.6%) over 40 years of age. No clinical or epidemiological differences were found between Paracoccidioides species. However, it is important to note that the only case of P. lutzii died as an outcome. CONCLUSIONS: This study suggests P. brasiliensis sensu stricto (S1) as the predominant species, showing its wide geographic distribution in Brazil. Furthermore, our findings revealed, for the first time, the occurrence of P. restrepiensis (PS3) in the state of Mato Grosso do Sul, Brazil. Despite our setbacks, it would be interesting to provide the complete sequencing of these clinical isolates to complement the molecular information presented.

Promoter-specific expression of the imprinted IGF2 gene in bovine oocytes and pre-implantation embryos.

The bovine IGF2 locus is a genomic region with alternative transcripts controlled by five promoters (P0, P1, P2, P3 and P4). As transcriptional regulation can affect messenger RNA (mRNA) stability and translation, and thus, subsequent biological effects, this study evaluated the bovine IGF2 promoter-specific expression patterns in oocytes and pre-implantation embryos produced in vitro by our standard IVP procedures. Immature and matured oocytes, and pre-implantation embryos at the 1-, 2-, 4-, 8- and 16-cell, and at early morula, compact morula, blastocyst and expanded blastocyst stages were collected in three pools of five structures per stage, in four replicates. Total RNA was extracted and subjected to RT-qPCR, using four sets of IGF2 promoter-specific primers covering transcripts driven by promoters P0/P1, P2, P3 and P4, with fragments sequenced for confirmation. Expression of P2- and P4-derived transcripts showed an initial peak between immature (P4) or matured (P2/P4) oocytes and 2-cell embryos, gradually falling until embryo genome activation (EGA), rising again at compaction and cavitation. P0/P1-derived transcripts were identified after EGA, during compaction, whereas P3 activity was not detected at any stage. Our findings suggest that P0/P1 and P2 likely have secondary roles during early stages, whereas P3 may be more relevant later in development. P4 seems to be the main pathway for bovine IGF2 expression during oocyte maturation and embryo development and, therefore, the main target to influence IVP in modulation of embryo growth and in studies in developmental biology.

A retrospective cohort study to screen linezolid-induced thrombocytopenia in adult patients hospitalized in the Midwestern Region of Brazil

ABSTRACT Background: Thrombocytopenia (TP) is the major event associated with linezolid (LZD) therapy. We investigated the incidence and risk factors for thrombocytopenia in hospitalized adults who received LZD (1200 mg/day) between 2015 and 2017. HIV-positive, death during follow-up and those with a baseline platelet count ≤100 × 103/mm3 were excluded. Method: TP was defined as a decrease in platelet count of ≥20% from the baseline level at the initiation of linezolid therapy and a final count of <100 × 103/mm3. The odds ratios (OR) for thrombocytopenia were obtained using multivariate stepwise logistic regression analysis. Main results: A total of 66 patients were included (mean age [SD] 62 [18], male gender [%], 37 [56]). LZD-associated TP was identified in 12 patients (18.2%). For TP, the adjusted OR [95% CI] of the platelet count ≤200 × 103/mm3, serum creatinine and renal impairment at baseline were 5.66 [1.15-27.9], 4.57 [1.26-16.5] and 9.41 [1.09-80.54], respectively. Male gender and dosage per weight per day (DPWD) >20 mg/kg/day were not risk factors. Conclusion: The results showed that the incidence of linezolid-induced thrombocytopenia was lower in patients with normal renal function and higher in those with platelet counts ≤200 × 103/mm3 or serum creatinine >1.5 mg/dL at the start of the treatment.

In vitro development of IVF-derived bovine embryos following cytoplasmic microinjection for the episomal expression of the IGF2 gene.

Important genomic imprinting changes usually occur following the in vitro production (IVP) of bovine embryos, especially in the imprinting pattern of components of the IGF system. This study aimed to evaluate the effects of a transient episomal overexpression of the IGF2 gene in bovine IVP embryos following embryo cytoplasmic microinjection (CMI) at the 1-cell stage on embryo survival, early and late developmental kinetics and morphological quality up to Day 7 of development. Selected cumulus-oocyte complexes (COCs) were matured and fertilized in vitro and subsequently segregated into six experimental groups: non-CMI control group and five CMI groups at increasing doses (0, 10, 20, 40 and 80 ng/µl) of a GFP vector built for the episomal expression of bovine IGF2. Zygote CMI was effective in delivering the expression vector into the ooplasm, irrespective of the groups, with 58% of positive GFP fluorescence in Day 7 blastocysts. Considering developmental rates and late embryo kinetics, the 10-ng/µl CMI vector dose promoted a lower blastocyst rate (10.4%), but for blastocysts at more advanced stages of development (93.0% blastocysts and expanded blastocysts), and higher number of cells (116.0 ± 3.0) than non-CMI controls (23.3%, 75.0% and 75.0 ± 6.8 were obtained, respectively). In conclusion, CMI at the 1-cell stage did not compromise subsequent in vitro development of surviving embryos, with the 10-ng/µl group demonstrating a possible growth-promoting effect of the IGF2 gene on embryo development, from the 1-cell to the blastocyst stage.

A retrospective cohort study to screen linezolid-induced thrombocytopenia in adult patients hospitalized in the Midwestern Region of Brazil.

BACKGROUND: Thrombocytopenia (TP) is the major event associated with linezolid (LZD) therapy. We investigated the incidence and risk factors for thrombocytopenia in hospitalized adults who received LZD (1200mg/day) between 2015 and 2017. HIV-positive, death during follow-up and those with a baseline platelet count ≤100×103/mm3 were excluded. METHOD: TP was defined as a decrease in platelet count of ≥20% from the baseline level at the initiation of linezolid therapy and a final count of <100×103/mm3. The odds ratios (OR) for thrombocytopenia were obtained using multivariate stepwise logistic regression analysis. MAIN RESULTS: A total of 66 patients were included (mean age [SD] 62 [18], male gender [%], 37 [56]). LZD-associated TP was identified in 12 patients (18.2%). For TP, the adjusted OR [95% CI] of the platelet count ≤200×103/mm3, serum creatinine and renal impairment at baseline were 5.66 [1.15-27.9], 4.57 [1.26-16.5] and 9.41 [1.09-80.54], respectively. Male gender and dosage per weight per day (DPWD) >20mg/kg/day were not risk factors. CONCLUSION: The results showed that the incidence of linezolid-induced thrombocytopenia was lower in patients with normal renal function and higher in those with platelet counts ≤200×103/mm3 or serum creatinine >1.5mg/dL at the start of the treatment.

Overview of artemisinin effectiveness during outset years of its implementation in the western Brazilian Amazon.

BACKGROUND: The elimination of malaria depends on the blocking of transmission and of an effective treatment. In Brazil, artemisinin therapy was introduced in 1991, and here we present a performance overview during implementation outset years. METHODS: It is a retrospective cohort (1991 to 2002) of patients treated in a tertiary centre of Manaus, with positive microscopic diagnosis of Plasmodium falciparum malaria, under treatment with using injectable or rectal artemisinin derivatives, and followed over 35-days to evaluate parasite clearance, death and recurrence. FINDINGS: This cohort outcome resulted 97.6% (1554/1593) of patients who completed the 35-day follow-up, 0.6% (10/1593) of death and 1.8% (29/1593) of follow-up loss. All patients that died and those that presented parasitaemia recurrence had pure P. falciparum infections and received monotherapy. Considering patients who completed 35-day treatment, 98.2% (1527/1554) presented asexual parasitaemia clearance until D4 and 1.8% (27/1554) between D5-D10. It is important to highlight that had no correlation between the five treatment schemes and the sexual parasite clearance. Finally, it is noteworthy that we were able to observe also gametocytes carriage during all follow-up (D0-D35). MAIN CONCLUSIONS: Artemisinin derivatives remained effective in the treatment of falciparum malaria during first 12-years of use in north area of Brazil.

Overview of artemisinin effectiveness during outset years of its implementation in the western Brazilian Amazon

BACKGROUND The elimination of malaria depends on the blocking of transmission and of an effective treatment. In Brazil, artemisinin therapy was introduced in 1991, and here we present a performance overview during implementation outset years. METHODS It is a retrospective cohort (1991 to 2002) of patients treated in a tertiary centre of Manaus, with positive microscopic diagnosis of Plasmodium falciparum malaria, under treatment with using injectable or rectal artemisinin derivatives, and followed over 35-days to evaluate parasite clearance, death and recurrence. FINDINGS This cohort outcome resulted 97.6% (1554/1593) of patients who completed the 35-day follow-up, 0.6% (10/1593) of death and 1.8% (29/1593) of follow-up loss. All patients that died and those that presented parasitaemia recurrence had pure P. falciparum infections and received monotherapy. Considering patients who completed 35-day treatment, 98.2% (1527/1554) presented asexual parasitaemia clearance until D4 and 1.8% (27/1554) between D5-D10. It is important to highlight that had no correlation between the five treatment schemes and the sexual parasite clearance. Finally, it is noteworthy that we were able to observe also gametocytes carriage during all follow-up (D0-D35). MAIN CONCLUSIONS Artemisinin derivatives remained effective in the treatment of falciparum malaria during first 12-years of use in north area of Brazil.

Clinical and laboratorial features of oral candidiasis in HIV-positive patients.

INTRODUCTION: We describe the clinical and laboratorial features of oral candidiasis in 66 HIV-positive patients. METHODS: Polymerase chain reaction-based techniques were performed for differentiation of Candida spp. isolated from patients at a public teaching hospital in Midwest Brazil. RESULTS: Oral lesions, mainly pseudomembranous, were significantly related to higher levels of immunosuppression. Of 45 Candida isolates, 66.7% were C. albicans. Most of the isolates were susceptible to the antifungal drugs tested. CONCLUSIONS: Oral lesions were associated with higher immunosuppression levels. Lower susceptibility to antifungals by non-albicans isolates supports the importance of surveillance studies using susceptibility tests to aid in the treatment.

Clinical and laboratorial features of oral candidiasis in HIV-positive patients

Abstract INTRODUCTION We describe the clinical and laboratorial features of oral candidiasis in 66 HIV-positive patients. METHODS: Polymerase chain reaction-based techniques were performed for differentiation of Candida spp. isolated from patients at a public teaching hospital in Midwest Brazil. RESULTS: Oral lesions, mainly pseudomembranous, were significantly related to higher levels of immunosuppression. Of 45 Candida isolates, 66.7% were C. albicans. Most of the isolates were susceptible to the antifungal drugs tested. CONCLUSIONS: Oral lesions were associated with higher immunosuppression levels. Lower susceptibility to antifungals by non-albicans isolates supports the importance of surveillance studies using susceptibility tests to aid in the treatment.

Variability in the clinical distributions of Candida species and the emergence of azole-resistant non-Candida albicans species in public hospitals in the Midwest region of Brazil

Abstract INTRODUCTION Incidence and antifungal susceptibility of Candida spp. from two teaching public hospitals are described. METHODS The minimum inhibitory concentrations of fluconazole, voriconazole, itraconazole, and amphotericin B were determined using Clinical Laboratory Standard Institute broth microdilution and genomic differentiation using PCR. RESULTS Of 221 Candida isolates, 50.2% were obtained from intensive care unit patients; 71.5% were recovered from urine and 9.1% from bloodstream samples. Candida parapsilosis sensu stricto was the most common candidemia agent. CONCLUSIONS We observed variations in Candida species distribution in hospitals in the same geographic region and documented the emergence of non-C. albicans species resistant to azoles.

Osteología del miembro torácico de la corzuela parda Mazama gouazoubira (g. Fischer, 1814) (Cetartiodactyla: Cervidae) / Osteology of the thoracic limb of south american gray brocket Mazama gouazoubira (Fischer, 1814) (Cetartiodactyla: Cervidae)

El objetivo del estudio fue describir las características anatómicas y establecer hipótesis morfo-funcionales del esqueleto del miembro torácico de Mazama gouazoubira. Para ello, se utilizaron los miembros torácicos de cuatro adultos jóvenes. Los huesos se prepararon mediante ebullición y fueron blanqueados en una solución de peróxido de hidrógeno para obtener descripciones comparativas, mediciones osteométricas y radiografías digitales. En comparación con los rumiantes domésticos, la escápula resultó ser ancha y plana, con el acromion pequeño, el proceso coracoides muy pequeño y sin tuberosidad de la espina. El húmero se observó alargado con la diáfisis redondeada en una sección transversal y con proyecciones no articulares lisas. La ulna se unió al radio hasta la epífisis distal y juntos formaron un espacio interóseo proximal alargado y estrecho, sin surco vascular. El esqueleto de la mano presentó los metacarpianos III y IV bien desarrollados y fusionados, mientras que los metacarpianos II y V resultaron rudimentarios con disposición telemetacarpiana, filogenéticamente típico de cérvidos del Nuevo Mundo. Se encontraron cuatro dedos con tres falanges en cada uno, de los cuales, dos corresponden a los principales (III y IV) que llegan al suelo y dos son rudimentarios (II y V). Las radiografías nos permitieron visualizar los patrones de tensiones trabeculares normales y la osteometría permitió establecer relaciones con fines comparativos. Se reconocieron las adaptaciones esqueléticas del miembro torácico para favorecer la locomoción cursorial saltatoria.

This study aimed to describe anatomical features of the forelimb skeleton of Mazama gouazoubira in order to establish morphofunctional activity. Forelimbs of four young adult specimens were used to this end. The bones were prepared by boiling and cleared in a solution of hydrogen peroxide for comparative descriptions, osteometrics measurements and digital radiographs. Compared to domestic ruminants, the scapula proved to be wide and flat, with a small acromion, reduced coracoid process and lacking of spine tuberosity. The humerus had an elongated shaft with a rounded cross-section and discrete non-articular projections. The ulna accompanied the radio to the distal epiphysis and formed a long and narrow, proximally placed interosseous space without a vascular groove. The skeleton of the hand presented the rudimentary metacarpals II and V and the well-developed III and IV fused metacarpals, which is a typical telemetacarpal arrangement of close phylogenetically deer species from the New World. There were four fingers, each one with three phalanges; two main (III and IV) touching the ground and two rudimentary (II and V). Radiographs allowed visualizing patterns of normal trabecular tensions and osteometrics enabled to establish ratios for comparative purposes. Adaptations of skeletal forelimb to favor cursorial saltatory locomotion were typically identified.

Variability in the clinical distributions of Candida species and the emergence of azole-resistant non-Candida albicans species in public hospitals in the Midwest region of Brazil.

INTRODUCTION: Incidence and antifungal susceptibility of Candida spp. from two teaching public hospitals are described. METHODS: The minimum inhibitory concentrations of fluconazole, voriconazole, itraconazole, and amphotericin B were determined using Clinical Laboratory Standard Institute broth microdilution and genomic differentiation using PCR. RESULTS: Of 221 Candida isolates, 50.2% were obtained from intensive care unit patients; 71.5% were recovered from urine and 9.1% from bloodstream samples. Candida parapsilosis sensu stricto was the most common candidemia agent. CONCLUSIONS: We observed variations in Candida species distribution in hospitals in the same geographic region and documented the emergence of non-C. albicans species resistant to azoles.

Brachial Plexus in the Pampas Fox (Lycalopex gymnocercus): a Descriptive and Comparative Analysis.

Twenty thoracic limbs of ten Lycalopex gymnocercus were dissected to describe origin and distribution of the nerves forming brachial plexuses. The brachial plexus resulted from the connections between the ventral branches of the last three cervical nerves (C6, C7, and C8) and first thoracic nerve (T1). These branches connected the suprascapular, subscapular, axillary, musculocutaneous, radial, median and ulnar nerves to the intrinsic musculature and connected the brachiocephalic, thoracodorsal, lateral thoracic, long thoracic, cranial pectoral and caudal pectoral nerves to the extrinsic musculature. The C7 ventral branches contribute most to the formation of the nerves (62.7%), followed by C8 (58.8%), T1 (40.0%) and C6 (24.6%). Of the 260 nerves dissected, 69.2% resulted from a combination of two or three branches, while only 30.8% originated from a single branch. The origin and innervation area of the pampas fox brachial plexus, in comparison with other domestic and wild species, were most similar to the domestic dog and wild canids from the neotropics. The results of this study can serve as a base for comparative morphofunctional analysis involving this species and development of nerve block techniques. Anat Rec, 300:537-548, 2017. © 2016 Wiley Periodicals, Inc.

Morfologia da laringe em Cerdocyon thous (Linnaeus, 1766) / Laryngeal morphology in Cerdocyon thous (Linnaeus, 1766)

Cerdocyon thous (cachorro-do-mato) é o canídeo silvestre mais comum em território sul-americano. Objetivou-se com o presente trabalho descrever a morfologia macroscópica, microscópica e comparada da laringe nesta espécie. Para tal, as laringes de dezesseis espécimes (quatro machos e doze fêmeas) foram analisadas quanto à topografia, forma, morfometria, musculatura intrínseca e histologia. A laringe dispôs-se ventralmente ao áxis e foi constituída por uma cartilagem tireoide e outra cricoide (hialinas), uma epiglótica (elástica) e um par de ariteoides (mistas). Um par de cartilagens sesamoides foi identificado entre os processos corniculados e a lâmina da cricoide. A morfometria revelou que a tireoide é a maior cartilagem. Não houve sinais definitivos de dimorfismo sexual na laringe de C. thous. O epitélio predominante foi do tipo pavimentoso estratificado o qual sofreu transição para pseudoestratificado cilíndrico ciliado ao nível do terço caudal da tireoide e rostral da cricoide. A laringe de C. thous mostrou semelhança com a do cão doméstico, ainda que o formato das cartilagens tenha apresentado diferenças.

Cerdocyon thous (crab-eating fox) is the most common wild canid in South American territory. The aim of this paper was to describe macroscopic, microscopic and comparative morphology of the larynx of this species. Analyses of larynges of sixteen specimens (four males and twelve females) revealed the topography, conformation, morphometrics, intrinsic muscles and histological characteristics. The larynx was positioned ventral to the axis and comprised one cricoid and one thyroid cartilage (hyaline), one epiglottis (elastic) and a pair of arytenoids (mixed). A pair of sesamoid cartilages was identified between the corniculate processes and the cricoid lamina. Morphometry revealed that thyroid cartilage was the largest. There were no conclusive signs of sexual dimorphism in the larynx of this species. Squamous stratified epithelium predominated and underwent transition to ciliated pseudo-stratified at the level of caudal thyroid and rostral cricoid portions. By comparison, larynx of C. thous showed similarity with the domestic dog, although the shape of cartilages has shown some variation.

Morfologia da laringe em Cerdocyon thous (Linnaeus, 1766) / Laryngeal morphology in Cerdocyon thous (Linnaeus, 1766)

Cerdocyon thous (cachorro-do-mato) é o canídeo silvestre mais comum em território sul-americano. Objetivou-se com o presente trabalho descrever a morfologia macroscópica, microscópica e comparada da laringe nesta espécie. Para tal, as laringes de dezesseis espécimes (quatro machos e doze fêmeas) foram analisadas quanto à topografia, forma, morfometria, musculatura intrínseca e histologia. A laringe dispôs-se ventralmente ao áxis e foi constituída por uma cartilagem tireoide e outra cricoide (hialinas), uma epiglótica (elástica) e um par de ariteoides (mistas). Um par de cartilagens sesamoides foi identificado entre os processos corniculados e a lâmina da cricoide. A morfometria revelou que a tireoide é a maior cartilagem. Não houve sinais definitivos de dimorfismo sexual na laringe de C. thous. O epitélio predominante foi do tipo pavimentoso estratificado o qual sofreu transição para pseudoestratificado cilíndrico ciliado ao nível do terço caudal da tireoide e rostral da cricoide. A laringe de C. thous mostrou semelhança com a do cão doméstico, ainda que o formato das cartilagens tenha apresentado diferenças.(AU)

Cerdocyon thous (crab-eating fox) is the most common wild canid in South American territory. The aim of this paper was to describe macroscopic, microscopic and comparative morphology of the larynx of this species. Analyses of larynges of sixteen specimens (four males and twelve females) revealed the topography, conformation, morphometrics, intrinsic muscles and histological characteristics. The larynx was positioned ventral to the axis and comprised one cricoid and one thyroid cartilage (hyaline), one epiglottis (elastic) and a pair of arytenoids (mixed). A pair of sesamoid cartilages was identified between the corniculate processes and the cricoid lamina. Morphometry revealed that thyroid cartilage was the largest. There were no conclusive signs of sexual dimorphism in the larynx of this species. Squamous stratified epithelium predominated and underwent transition to ciliated pseudo-stratified at the level of caudal thyroid and rostral cricoid portions. By comparison, larynx of C. thous showed similarity with the domestic dog, although the shape of cartilages has shown some variation.(AU)

Origens e ramificações do plexo braquial no cachorro-do-mato Cerdocyon thous (Linnaeus, 1766) / Origins and ramifications of brachial plexus in the crab-eating-fox Cerdocyon thous (Linnaeus, 1766)

Este estudo teve o objetivo de descrever a origem e ramificação dos nervos de vinte plexos braquiais de cachorro-do-mato (Cerdocyon thous). Dez animais da espécie, obtidos post mortem por atropelamento em rodovias, foram utilizados para o estudo, de acordo com a autorização do IBAMA/SISBIO nº33667-1. Depois de coletados, os cadáveres foram fixados em solução de formaldeído a 50% e conservados por pelo menos 14 dias em solução de formaldeído a 10% antes das dissecções. Após remoção da pele, incisões na musculatura peitoral e afastamento dos membros, acessou-se o espaço axilar e os nervos do plexo tiveram seu trajeto dissecados individualmente até sua inserção muscular. Para melhorar a visualização dos ramos ventrais cervicais e torácicos que originavam cada nervo, removeram-se a musculatura que encobria os forames intervertebrais, os processos transversos e os corpos vertebrais ventralmente, expondo inclusive a medula espinhal. Registros fotográficos e desenhos esquemáticos documentaram a origem e ramificação dos nervos. Os vinte plexos braquiais foram resultantes das conexões entre os ramos ventrais dos três últimos nervos espinhais cervicais (C6, C7 e C8) e do primeiro torácico (T1). Estes ramos derivaram os nervos supra-escapular, subescapular, axilar, musculocutâneo, radial, mediano, ulnar para a musculatura intrínseca e os nervos braquiocefálico, toracodorsal, torácico lateral, torácico longo, peitoral cranial e peitoral caudal para a musculatura extrínseca do membro torácico. Constatou-se que os ramos ventrais de C7 foram os que mais contribuíram na formação de nervos (61,5%), seguido de C8 (55,4%), de T1 (41,2%) e de C6 (30,8%). O teste t de comparação entre as médias, ao nível de significância de 5%, não demonstrou diferenças na origem do plexo quando comparados os antímeros e os sexos. Do total dos 260 nervos dissecados, 68,8% foram originados pela combinação de dois ou três ramos, enquanto apenas 31,2% tiveram formação por um único ramo. A combinação entre C8 e T1 foi a que mais formou nervos para o plexo (23,8%) nesta espécie. Quando comparadas a origem, ramificação e área de inervação do plexo braquial do C. thous com outras espécies domésticas e silvestres, verificou-se maior semelhança com o cão doméstico. Os resultados deste estudo podem embasar o diagnóstico de disfunções neuromusculares, as técnicas de bloqueios anestésicos e análises morfofuncionais comparativas envolvendo esta espécie.(AU)

This study aimed to describe the origin and branching of nerves from twenty brachial plexuses of crab-eating foxes (Cerdocyon thous). Ten animals of the species, obtained post mortem from being run over on highways, were used for the study, in accordance with the authorization from IBAMA / SISBIO No. 33667-1. Once collected, the cadavers were fixed in 50% formaldehyde and kept for at least 14 days in a solution of 10% formaldehyde before dissections. After removal of skin, incisions in breast muscles and reflection of thoracic limbs allowed access to axillary space and the nerves could have trajects dissected individually to each muscle insertion. To improve visualization of the cervical and thoracic ventral roots that originated every single nerve, muscles that covered the intervertebral foramina, transverse processes and vertebral bodies were removed ventrally and the spinal cord exposed. Schematic drawings and photographic records documented the origin and branching of nerves. The twenty plexuses were resulted from connections between the ventral branches of the last three cervical spinal nerves (C6, C7 and C8) and first thoracic (T1). These branches derived the nerves suprascapular, subscapular, axillary, musculocutaneous, radial, median and ulnar to the intrinsic muscles and brachiocephalic, thoracodorsal, lateral thoracic, long thoracic, cranial and caudal pectoral nerves to the extrinsic muscles of the thoracic limb. It was found that the ventral rami of C7 were the main contributors in the formation of nerves (61.5%), followed by C8 (55.4%), T1 (41.2%) and C6 (30.8%). The t-test comparison between means at a significance level of 5% showed no differences in the origin of plexus when compared antimeres and sexes. Of the total of 260 dissected nerves, 68.8% originated by the combination of two or three roots, while only 31.2% were formed by a single root. The combination between C8 and T1 was the most frequent origin of nerves to the plexus (23.8%) in this species. Comparing the origin, branching and innervation area of the brachial plexus in C. thous with other domestic and wild species, there was a greater similarity with the domestic dog. These results may give the anatomical basis to diagnosis of neuromuscular disorders, anesthetic blocks techniques and comparative morphofunctional analyzes involving this species.(AU)