Hepatic transcriptome profiling of largemouth bass (Micropterus salmoides Lacépède) injected with Flavobacterium covae or lipopolysaccharide.

Flavobacterium covae (columnaris) is the most detrimental bacterial disease affecting the largemouth bass (Micropterus salmoides Lacépède) aquaculture industry. In the current study, fish received an intraperitoneal injection of either 1× PBS (100 µL), LPS in PBS (100 µL, 10 µg/mL), or F. covae (100 µL, 2.85 × 1011 CFU/mL) to simulate immunological challenges. After 24 h post-injection, liver tissue from the control and treated groups were then collected for transcriptome analysis. Results of the Gene Ontology (GO) and KEGG pathway analyses for the F. covae and LPS-injected groups found differentially expressed genes (DEGs) enriched primarily in toll-like receptors (TLRs), cytokine-cytokine receptors, complement and coagulation cascades, and the PPAR signalling pathways. This suggests that the liver immune system is enhanced by these five combined pathways. Additionally, the DEGs TLR5, MYD88, and IL-1 were significantly upregulated in F. covae and LPS-injected fish compared to the controls, whereas IL-8 was downregulated. The upregulation of TLR5 was unexpected as F. covae lacks flagellin, the protein that binds to TLR5. Additionally, it is unknown whether the TLR5 is upregulated by LPS. Further research into the upregulation of TLR5 is warranted. These results provide insight into immune responses and associated pathways contributing to the immune system in the liver during columnaris infection and induced response to LPS in largemouth bass.

Extracorporeal Testicular Ectopia through Femoral Region: A Rare Variant of Scrotoschisis.

A 2-year-old male child presented to us with absent left testis in scrotum since birth. Parents noted a pinkish white globular mass in medial aspect of left thigh. At the time of presentation (2 years old) he had a 2 x 2 cm, firm, subcutaneous swelling located on the medial aspect of the left thigh. Ultrasonography was suggestive of left ectopic testis of size 1 x 1.2cm in the femoral region. Orchidopexy was done. In our case the term scrotoschisis will not be appropriate as testicular extrusion has occurred through femoral region and not the scrotum. This can be considered as extracorporeal testicular ectopia or bubonoschisis, respective to the location of the defect. We are reporting a case of extra corporeal testicular ectopia through the femoral region for the first time.

Neonatal Sigmoid Colon Perforation in Low Anorectal Malformation.

Anorectal malformations (ARMs) are common congenital anomalies in neonates. Colonic perforation is very rare in ARMs. Delay in diagnosis of neonate with ARM results in colonic perforations and life-threatening morbidity. Colonic perforation due to ARM may not be completely avoided; however, early diagnosis and management are essential in assuring better outcomes.

Different dietary combinations of high/low starch and fat with or without bile acid supplementation on growth, liver histopathology, gene expression and fatty acid composition of largemouth bass, Micropterus salmoides.

High dietary levels of fat and/or starch can lower the growth and cause extensive liver inflammation that is linked to mortalities in largemouth bass, Micropterus salmoides. However, bile acids (BA) may mitigate these adverse effects. In a 2 × 2 × 2 factorial feeding trial, M. salmoides juveniles were fed different combinations of dietary high (HF), low fat (LF), high (HS) or low starch (LS) levels with or without BA supplementations at 1% for 8 weeks. A total of 8 isonitrogenous diets were formulated to include, HF/LS, HF/HS, LF/HS, LF/LS, HF/LS-BA, HF/HS-BA, LF/HS-BA and LF/LS-BA. Survival, growth performance, feeding efficiency, whole-body proximate composition, muscle/liver fatty acid composition, hepatic expression of growth regulator (GH/IGF1 axis), lipid metabolism (fatty acid synthase 'FASN' and cholesterol 7 alpha-hydroxylase 'CYP7A1') and antioxidant capacity (superoxide dismutase 'SOD') genes as well as liver histopathology were assessed. Results showed that among diets without BA, there was no significant effect on growth or feeding efficiency, but when BA was included this led to more variable effects including significantly higher weight gain in the LF/HS-BA group compared to all others fed BA. The HF, HS or their combination led to extensive hepatic inflammation, but BA appeared to partially mitigate this in the LF/HS group (i.e. LF/HS-BA). No abnormal liver histopathology was observed in the LF/LS and LF/LS-BA treatments. Muscle 22:6n-3 was significantly higher in the HF/LS and HF/HS-BA groups compared to those fed the HF/HS or LF/LS diets. Dietary fat had a significant effect on the moisture, crude lipid, and caloric content of M. salmoides. Hepatic expression of IGF-I and CYP7A1 were differentially modulated under different treatments. Overall, these results show that BA can alleviate some liver inflammation caused by high dietary starch; however the LF/LS diets led to a better balance between growth performance and liver health.

Chlorpyrifos and dichlorvos in combined exposure reveals antagonistic interaction to the freshwater fish Mrigal, Cirrhinus mrigala.

Toxicity imposed by organophosphate pesticides to the freshwater cultivable fish species mrigal (Cirrhinus mrigala) was assessed under laboratory conditions. Healthy juveniles were exposed to chlorpyrifos, dichlorvos, and their equitoxic mixture in geometric series. Median lethal concentrations of chlorpyrifos were found to be 0.906 (0.689-1.179), 0.527 (0.433-0.633), 0.435 (0.366-0.517) and 0.380 (0.319-0.450) mg/L and dichlorvos were found to be 38.432 (33.625-47.866), 22.477 (19.047-26.646), 12.442 (9.619-14.196) and 11.367 (9.496-13.536) mg/L after 24 h, 48 h, 72 h and 96 h of exposure respectively. Surprisingly, the joint toxicity of these organophosphates in the binary mixture was less than additive during most of the exposure periods. Behavioral changes exhibited by individual as well as mixture pesticide treatments were loss of schooling behavior, aggregating at corners of the test chamber, elevated opercular beatings, surplus mucus secretion, slight color changes and sudden and rapid body movements before death. Loss of fish equilibrium was noticed only in chlorpyrifos treated fish, whereas sluggish behavior was noticed only in mixture pesticide treatment. Such behavioral studies can be applied as a non-invasive bio-monitoring tool for water quality assessment for fish growth and development. Despite the same mode of action of both pesticides, the antagonistic action in the binary mixture is an interesting outcome of this research that requires further investigation for a lucid understanding of the joint toxicity mechanism of such pesticides.

Colonic Agenesis with Anorectal Malformation - Rarest Variant of Congenital Short Colon.

We report a case of colonic agenesis with anorectal malformation in a newborn girl. The patient also presented with congenital heart disease. We are presenting the clinical features, intraoperative findings, and treatment plan.

Coping strategies in response to different levels of elevated water hardness in channel catfish (Ictalurus punctatus): Insight into ion-regulatory and histopathological modulations.

Water hardness above the optimal level can incite toxic effects in fish, which are often species specific. Hence, we aimed at obtaining insights on the potential effects of elevated water hardness as well as coping strategies in channel catfish (Ictalurus punctatus). First, a toxicity assay was performed where the 96 h-LC50 was calculated as 4939 mg/L CaCO3. Thereafter, to gain knowledge on the underlying adaptive strategies to high water hardness, fish were exposed to seven hardness levels (150, 600, 1000, 1500, 2000, 3000 and 4000 mg/L CaCO3 at pH 8.15) for 15 days. Results showed that branchial activities of Ca2+-ATPase and Na+/K+-ATPase, which facilitate Ca2+ uptake, reduced starting respectively from 1000 mg/L and 1500 mg/L CaCO3. Nevertheless, Ca2+ burden in plasma and tissue (gills, liver and intestine) remained elevated. Hardness exposure also disturbed cations (Na+, K+, Mg2+) and minerals (iron and phosphorus) homeostasis in a tissue-specific and dose-dependent manner. Both hemoglobin content and hematocrit dropped significantly at 3000-4000 mg/L CaCO3, with a parallel decline in iron content in plasma and gills. Muscle water content rose dramatically at 4000 mg/L CaCO3, indicating an osmo-regulation disruption. Higher hardness of 3000-4000 mg/L CaCO3 also incited a series of histopathological modifications in gills, liver and intestine; most likely due to excess Ca2+ accumulation. Overall, these data suggest that channel catfish can adapt to a wide range of elevated hardness by modulating Ca2+ regulatory pathways and histomorphological alterations, however, 1500 mg/L CaCO3 and above can impair the performance of this species.

Effects of bioprocessed soybean meal and nucleotide supplementation on growth, physiology and histomorphology in largemouth bass, Micropterus salmoides, juveniles.

In this 8-week feeding trial, the effects of nucleotide (N) supplementation (at 0.05%) were compared in diets with conventional soybean meal (CSBM or CSBM + N) versus bioprocessed SBM (BSBM or BSMB + N) on largemouth bass, Micropterus salmoides, juveniles. A total of five isonitrogenous and isolipidic diets were formulated, with the control diet being fishmeal-based. Growth, feeding efficiency, proximate composition, hepatic expression of genes involved in lipid metabolism and growth as well as liver/intestinal histopathology were assessed. Results showed that growth was significantly higher in fish fed the control diet, but there was no significant effect of SBM type or nucleotide supplementation on growth, feeding efficiency, or proximate composition. Hepatic expression of growth hormone (GH), insulin-like growth factor I (IGFI), superoxide dismutase (SOD), fatty acid synthase (FASN) and cholesterol 7 alpha-hydroxylase (CYP7A1) were unaffected by the diets. Tumor necrosis factor alpha (TNF-α) and transforming growth factor beta (TGF-ß) were significantly downregulated and upregulated, respectively, in the SBM-based treatments compared with the control. The intestinal villi were significantly shorter and wider in fish fed the CSBM diet compared to the other treatments. The villi height and width were similar between the control and those fed the BSMB + N diet. It may be possible that the unaffected growth by nucleotides were due to an insufficient dose and/or undisrupted nucleotide synthesis due to being cultured under good conditions. Meanwhile, the unaffected growth in the SBM treatment could indicate a tolerance of M. salmoides to plant proteins and associated antinutritional factors. Nevertheless, BSBM and/or nucleotides appeared to mitigate some adverse effects of dietary SBM to the intestinal histomorphology in M. salmoides.

Physio-biochemical, metabolic nitrogen excretion and ion-regulatory assessment in largemouth bass (Micropterus salmoides) following exposure to high environmental iron.

Iron overload is a significant water quality issue in many parts of the world. Therefore, we evaluated the potential toxic effects of waterborne elevated iron on largemouth bass (Micropterus salmoides), a highly valued sport and aquaculture fish species. First, a 96 h-LC50 toxicity assay was performed to understand the tolerance limit of this species to iron; and was determined to be 22.07 mg/L (as Fe3+). Thereafter, to get a better insight on the fish survival during long-term exposure to high environmental iron (HEI) (5.52 mg/L, 25% of the determined 96 h-LC50 value), a suite of physio-biochemical, nitrogenous metabolic and ion-regulatory compensatory responses were examined at 7, 14, 21 and 28 days. Results showed that oxygen consumption dropped significantly at 21 and 28 days of HEI exposure. Ammonia excretion rate (Jamm) was significantly inhibited from day 14 and remained suppressed until the last exposure period. The transcript concentration of Rhesus glycoproteins Rhcg2 declined; likely diminishing ammonia efflux out of gills. These changes were also reflected by a parallel increment in plasma ammonia levels. Under HEI exposure, ion-balance was negatively affected, manifested by reduced plasma [Na+] and parallel inhibition in branchial Na+/K+-ATPase activity. Muscle water content was elevated in HEI-exposed fish, signifying an osmo-regulatory compromise. HEI exposure also increased iron burden in plasma and gills. The iron accumulation pattern in gills was significantly correlated with a suppression of Jamm, branchial Rhcg2 expression and Na+/K+-ATPase activity. There was also a decline in the glycogen, protein and lipid reserves in the hepatic tissue from 14 days, 28 days and 21 days, respectively. Overall, we conclude that sub-lethal chronic iron exposure can impair normal physio-biochemical and ion-regulatory functions in largemouth bass. Moreover, this data set can be applied in assessing the environmental risk posed by a waterborne iron overload on aquatic life.

Anorectal Malformations in Males: Pros and Cons of Neonatal versus Staged Reconstruction for High and Intermediate Varieties.

BACKGROUND: High and intermediate types of anorectal malformations (ARMs) in male neonates may be managed either by primary neonatal reconstruction without colostomy cover or by traditional policy of staged reconstruction after neonatal colostomy. Posterior sagittal anorectoplasty (PSARP) is the current widely practiced reconstructive technique with varied results. AIM: To assess our functional results of PSARP without colostomy in male neonates with high and intermediate ARMs compared to 3-stage (neonatal colostomy - PSARP - colostomy closure) methodology in a high volume tertiary care institution of a developing country. PATIENTS AND METHODS: The number of colostomies performed for male high/intermediate anomalies and the number of babies who completed 3-stage reconstruction during a 10-year period is analyzed. The outcome of primary neonatal PSARPs during the same period was analyzed. Eighty primary PSARPs were compared to 81 staged reconstructions for outcome analysis, using Kelly score. RESULTS: A total of 453 colostomies were performed, but only 253 of them completed all stages of reconstruction (52%). Good continence was achieved in 45% of cases of primary PSARP versus 26% in 3-staged surgery. CONCLUSIONS: Primary PSARP in neonatal period without colostomy is a good option for high and intermediate ARMs in males if the treating surgeon is reasonably skilled in neonatal surgery and PSARP procedures.

Currarino syndrome: Rare clinical variants.

Currarino syndrome (CS) is a rare clinical condition. The classical presentation includes a triad of sacral anomaly, anorectal malformations, and presacral mass. This syndrome belongs to the group of persistent neuroenteric malformations. This article presents two cases of Currarino syndrome, where there was rare clinical variants such as rectal atresia in the first case and rectal stenosis in the second case. The clinical presentations were very deceptive as the first case presented as high anorectal malformation and the second case was simulating Hirschprung's disease.

The combined effect of hypoxia and nutritional status on metabolic and ionoregulatory responses of common carp (Cyprinus carpio).

In the present study, the combined effects of hypoxia and nutritional status were examined in common carp (Cyprinus carpio), a relatively hypoxia tolerant cyprinid. Fish were either fed or fasted and were exposed to hypoxia (1.5-1.8mg O2L(-1)) at or slightly above their critical oxygen concentration during 1, 3 or 7days followed by a 7day recovery period. Ventilation initially increased during hypoxia, but fasted fish had lower ventilation frequencies than fed fish. In fed fish, ventilation returned to control levels during hypoxia, while in fasted fish recovery only occurred after reoxygenation. Due to this, C. carpio managed, at least in part, to maintain aerobic metabolism during hypoxia: muscle and plasma lactate levels remained relatively stable although they tended to be higher in fed fish (despite higher ventilation rates). However, during recovery, compensatory responses differed greatly between both feeding regimes: plasma lactate in fed fish increased with a simultaneous breakdown of liver glycogen indicating increased energy use, while fasted fish seemed to economize energy and recycle decreasing plasma lactate levels into increasing liver glycogen levels. Protein was used under both feeding regimes during hypoxia and subsequent recovery: protein levels reduced mainly in liver for fed fish and in muscle for fasted fish. Overall, nutritional status had a greater impact on energy reserves than the lack of oxygen with a lower hepatosomatic index and lower glycogen stores in fasted fish. Fasted fish transiently increased Na(+)/K(+)-ATPase activity under hypoxia, but in general ionoregulatory balance proved to be only slightly disturbed, showing that sufficient energy was left for ion regulation.

Hypo-osmotic stress-induced physiological and ion-osmoregulatory responses in European sea bass (Dicentrarchus labrax) are modulated differentially by nutritional status.

We investigated the impact of nutritional status on the physiological, metabolic and ion-osmoregulatory performance of European sea bass (Dicentrarchus labrax) when acclimated to seawater (32 ppt), brackish water (20 and 10 ppt) and hyposaline water (2.5 ppt) for 2 weeks. Following acclimation to different salinities, fish were either fed or fasted (unfed for 14 days). Plasma osmolality, [Na(+)], [Cl(-)] and muscle water content were severely altered in fasted fish acclimated to 10 and 2.5 ppt in comparison to normal seawater-acclimated fish, suggesting ion regulation and acid-base balance disturbances. In contrast to feed-deprived fish, fed fish were able to avoid osmotic perturbation more effectively. This was accompanied by an increase in Na(+)/K(+)-ATPase expression and activity, transitory activation of H(+)-ATPase (only at 2.5 ppt) and down-regulation of Na(+)/K(+)/2Cl(-) gene expression. Ammonia excretion rate was inhibited to a larger extent in fasted fish acclimated to low salinities while fed fish were able to excrete efficiently. Consequently, the build-up of ammonia in the plasma of fed fish was relatively lower. Energy stores, especially glycogen and lipid, dropped in the fasted fish at low salinities and progression towards the anaerobic metabolic pathway became evident by an increase in plasma lactate level. Overall, the results indicate no osmotic stress in both feeding treatments within the salinity range of 32 to 20 ppt. However, at lower salinities (10-2.5 ppt) feed deprivation tends to reduce physiological, metabolic, ion-osmo-regulatory and molecular compensatory mechanisms and thus limits the fish's abilities to adapt to a hypo-osmotic environment.

Tubulocystic enteric duplication: a unique entity.

Enteric duplication has cystic and tubular varieties. A male infant presented with a large cystic, well-demarcated mass in the right flank. On exploratory laparotomy, multiple cystic and tubular lesions were present adjacent to the mesenteric border of the small bowel along with malrotation of the small bowel. The tubule-cystic structure was excised along with the involved normal bowel segment and Ladd's procedure was performed. Histopathological evaluation revealed an intestinal duplication cyst. The occurrence of midgut malrotation and volvulus along with duplication is uncommon. The cyst's substantial size could have been an aetiological factor for malrotation and volvulus. The child's small bowel had adapted remarkably with time. This case highlights a new variant of duplication cysts.

Female Anorectal Malformation with Genitourinary Prolapse: A Rare Association.

ABSTRACT: Genitourinary prolapse in newborn females as an introital mass is an uncommon entity. The usual causative mechanisms are poor pelvic innervation, damage or pressure on pelvic musculature and ligaments etc. Different methods of reduction as treatment were proposed in the past. Apart from uncommon occurrence of genitourinary prolapse in newborns, its association with anorectal malformation is not reported in English literature after searching on PubMed and Google Scholar. We report three cases of genitourinary prolapse with anorectal malformation in newborn females where decompressing colostomy was curative for the condition reflecting increased intra-abdominal pressure as causative mechanism.

Advocating for Routine Post-natal Surveillance and Urgent Neonatal Inguinal Hernia Repair.

ABSTRACT: Inguinal hernias are more common in preterm and neonates and incidence of incarceration are reported to be more in the first 6 months of life. Strangulation follows incarceration and various incarcerated and strangulated contents having been reported in the sac. The fistulation of the herniated content through the scrotal skin is quite rare with only a few reported cases. We present the case of entero-scrotal fistula in a neonate managed with staged repair along with a brief review of the literature. A 27-days-old, full-term male presented with faecal discharge from the right scrotum. He had no tell-tale signs of obstructed hernia. A faecal fistulous opening was located in the right hemi-scrotum. The terminal ileum was seen as the content with an antimesenteric perforation, divided ileostomy and scrotal debridement was done and later ileo-ascending anastomosis was performed electively. Neonatal hernias should be considered an urgency and we advocate early surgery. The resource-limited setting and poor post-natal surveillance may have added to the worries. In our case, swelling and fistulation occurred in a very brief period of 36 h. We managed the child with a diversion stoma followed by ileo-ascending anastomosis later.