Chloride-induced increment in short-circuiting current of the turtle bladder. Effects of in-vivo acid-base state

Evidence for the participation of conductive and non-conductive (exchange) transmembrane anion pathways in the luminal acidification, alkalinization, and chloride-reabsorptive functions of the turtle bladder is provided from the pattern of Cl- -induced changes in transepithelial electrical parameters of isolated urinary bladders from three groups of donor turtles: control or post-absorptive turtles (those killed 5 days after feeding); acidotic turtles (NH4Cl-loaded); and alkalotic turtles (NaHCO3-loaded). The predominance of each of the three aforementioned transport functions as well as the response to Cl- -addition is altered by the in-vivo electrolyte balance of the turtle. In post-absorptive bladders, which are poised for acidification and Cl- reabsorption, the mucosal and serosal addition of Cl- to Na+-free, (HCO3- + CO2)-containing media increases the negative short-circuiting current (Isc). In acidotic bladders, which are poised for acidification but not Cl- reabsorption, mucosal Cl- addition has no effect on this Isc whereas serosal Cl- addition increases the negative Isc in a manner identical to that observed in the post-absorptive bladders. Alkalotic bladders do not possess an acidification function but instead are poised for Cl- reabsorption and cAMP-dependent electrogenic alkali secretion (positive Isc). In these bladders, serosal Cl- addition is without effect while mucosal Cl- addition produces transient changes in this positive Isc. It is found that these results can be replicated by a model of the turtle bladder in which transmembrane Cl- and HCO3- conductive and exchange paths mediate transepithelial acidification, alkalinization and Cl- reabsorption.     

In vivo environmental temperature and the in vitro pattern of luminal acidification in turtle bladders. Evidence for HCO3 ion reabsorption

In this study, it is shown how to transfer tared aliquots of (HCO3 + CO2)-containing luminal fluids directly into the mercury-sealed chamber of a modified Van Slyke apparatus and how to obtain direct as well as indirect manometric determinations of dissolved CO2 ([CO2]f) in each aliquot of such fluids. It is next shown that the pattern of in vitro luminal acidification in an isolated turtle bladder sac depends upon the prior in vivo ambient temperature to which the donor turtle had become adapted. Under in vivo conditions, the food intake, physical activity, and acid excretion of 32 degrees C-adapted turtles are greater than those of 21 degrees C or 26 degrees C-adapted turtles. Under in vitro conditions of incubating isolated bladder sacs (from 21, 26, and 32 degrees C turtles) in (HCO3 + CO2)-containing Ringer media at a single temperature (21 degrees C), the patterns of luminal acidification are as follows: (a) The rate of depletion of luminal [HCO3] is greatest in bladders from the 32 degrees C-adapted turtles. (b) Concomitant decreases in luminal [CO2]f, [HCO3], and pH (the 'CO2-decreasing patterns' of luminal acidification) develop in all bladders from 32 degrees C turtles, in half of those from 26 degrees C turtles, but in less than one-fifth of those from 21 degrees C-adapted turtles: and (c) a CO2-increasing pattern of luminal acidification is found in most of the bladders from 21 degrees C-adapted turtles. A postulated bicarbonate ion-reabsorbing pump is consistent with all of these patterns of luminal acidification.     

Host-parasite interface of the urinary bladder-inhabiting nematode Trichosomoides crassicauda: changes induced in the urothelium of infected rats

The urinary bladders of rats infected with Trichosomoides crassicauda were studied by light, scanning and transmission electron microscopy. The bladder epithelium of infected rats showed a diffuse, mild, flat hyperplasia, four to six cells thick. The anterior parts of adult female worms were embedded in tunnels within the hyperplastic epithelium with the posterior portions of the parasites lying free in the bladder lumen. The hyperplastic epithelial cells forming the inner layer of the tunnel wall, adjacent to the parasite, showed degenerative changes. These cells contained single or multiple pycnotic nuclei. Their cytoplasm was fibro-granular in appearance, with few distinct organelles, and the luminal surfaces of the cells were not limited by plasma membranes. Small numbers of granular cells, similar in appearance to mast cells, were seen in, and possibly crossing, the epithelium. There was no infection-related increase in the number or type of inflammatory cells in the lamina propria. Questions of interest emerging from this study relate to the nutrition of an adult nematode occupying an intra-epithelial location, and the absence of a significant chronic inflammatory response to the mature worm. It is suggested that the avascular hyperplastic epithelium of the bladder is an immunologically 'protected site' for the mature female T. crassicauda.     

Insulin cells are found in the main and accessory urinary bladders of the painted turtle, Chrysemys picta

Insulin has been localized immunocytochemically to cells in the main and accessory urinary bladders of the painted turtle, Chrysemys picta, and represents an unusual addition to the specturm of regulatory peptides associated with the urinary bladder. These stellate to fibroblastoid cells often possess neural-like processes and are similar in morphology to neurotensin cells found in Chrysemys and Pseudemys urinary bladders. Radioimmunoassay of 2M acetic acid extracts of bladder tissue indicate that the insulin concentration of accessory bladder is several-fold greater than main bladder but considerably lower than the insulin content of pancreas. Pieces of accessory bladder incubated in vitro exhibit a stable insulin release into the medium over 1 hour, but release is unaltered by known insulin secretagogues. It is tempting to postulate an endocrine or paracrine regulatory function for these cells, but at present their role in Chrysemys bladder function remains unknown.     

Physiological role for vanadate-inhibitable active H+ transport: a new model for distal urinary acidification

We have shown elsewhere that membranes isolated from the turtle bladder epithelium have both vanadate-resistant (VR) and vanadate-sensitive (VS) components of ATP-dependent H+ transport. VR appears to be due to a vacuolar-type H+ uniport while VS has properties of an H/K exchanger. In the present study we used bladders from chronically alkalotic turtles, which do not acidify urine, and found them to yield membranes which retain VR but lack VS transport. Thus VS occurs only in membranes from bladders secreting acid, implying that VS H+ transport (1) plays a physiological role in the secretion and (2) is a functional marker for the apical membrane of the acid-secreting cells. These results suggest a new model for distal urinary acidification.     

Wild Cane Toads (Rhinella marina) Expel Foreign Matter from the Coelom via the Urinary Bladder in Response to Internal Injury,Endoparasites and Disease

Dissections of >1,200 wild-caught cane toads (Rhinella marina) in tropical Australia confirm a laboratory report that anurans can expel foreign objects from the coelom by incorporating them into the urinary bladder. The foreign objects that we found inside bladders included a diverse array of items (e.g., grass seeds, twigs, insect prey, parasites), many of which may have entered the coelom via rupture of the gut wall. In some cases, the urinary bladder was fused to other organs including liver, fat bodies, ovaries, Bidder’s organs, lungs, mesentery, stomach wall, gall bladder, and the abdominal wall. Acanthocephalan parasites (of a range of developmental stages) were identified from the walls of the urinary bladders of three cane toads. This organ may play a significant role in destroying or excreting metazoan parasites, as well as inanimate objects.     

Eradication of Arizona hinshawii from artificially infected turtle eggs.

Turtle eggs, 24 h old, were infected with Arizona hinshawii and treated 48 h later with gentamicin sulfate (Garasol; Shering Corp., Allantown, N.J.) by pressure differential egg dip treatment to ascertain the concentration of this reagent required to eradicate this pathogen from eggs. Infected eggs treated with 1,000 or 1,500 micrograms of gentamicin per ml of dip solution eliminated detectable A. hinshawii from eggs as determined by testing shells and embryo-yolk homogenates of 12-day-old eggs and the gastrointestinal tracts, kidneys, livers and gall bladders, and yolks of 50-day-old embryos. Treated eggs produced hatchlings which did not excrete detectable A. hinshawii at 72 h or 30 days after hatching, nor was this organism recovered from the visceral organs of these hatchlings when necropsied 30 days after hatching. Bacteriological assays on infected nontreated eggs showed that greater than 70% of the eggs harbored A. hinshawii, and eggs in this group produced hatchlings which actively excreted and harbored A. hinshawii. Eggs not infected or treated also produced turtles which excreted and systemically carried A. hinshawii and Salmonella spp. though not at the same level as did the turtles produced from infected, nontreated eggs.     

Eradication of Arizona hinshawii from artificially infected turtle eggs

Turtle eggs, 24 h old, were infected with Arizona hinshawii and treated 48 h later with gentamicin sulfate (Garasol; Shering Corp., Allantown, N.J.) by pressure differential egg dip treatment to ascertain the concentration of this reagent required to eradicate this pathogen from eggs. Infected eggs treated with 1,000 or 1,500 micrograms of gentamicin per ml of dip solution eliminated detectable A. hinshawii from eggs as determined by testing shells and embryo-yolk homogenates of 12-day-old eggs and the gastrointestinal tracts, kidneys, livers and gall bladders, and yolks of 50-day-old embryos. Treated eggs produced hatchlings which did not excrete detectable A. hinshawii at 72 h or 30 days after hatching, nor was this organism recovered from the visceral organs of these hatchlings when necropsied 30 days after hatching. Bacteriological assays on infected nontreated eggs showed that greater than 70% of the eggs harbored A. hinshawii, and eggs in this group produced hatchlings which actively excreted and harbored A. hinshawii. Eggs not infected or treated also produced turtles which excreted and systemically carried A. hinshawii and Salmonella spp. though not at the same level as did the turtles produced from infected, nontreated eggs.     

H+/ATP stoichiometry of proton pump of turtle urinary bladder

Urinary acidification in the turtle urinary bladder is due to a reversible proton-translocating ATPase. To estimate the H+/ATP stoichiometry of this pump, we measured the delta G'ATP in the epithelial cells and the maximum e.m.f. generated by the pump. The latter is the maximal transepithelial electrochemical gradient for protons placed across the epithelium that is needed to nullify the rate of transport and averaged 179 +/- 7 mV. The delta G'ATP averaged 50.1 kJ/mol. The H+/ATP stoichiometry of these bladders was 2.92 +/- 0.1. In other experiments, the bladders were poisoned by iodoacetate and cyanide and a variable transepithelial electrochemical gradient for protons was placed across them. It was noted that ATP synthesis occurred at a transepithelial electrochemical gradient for protons greater than 120 mV. The delta G'ATP in other bladders treated identically averaged 40.0 kJ/mol, giving a H+/ATP stoichiometry of 3.4 +/- 0.1. We conclude that the H+/ATP stoichiometry of the proton pump of turtle urinary bladder is approximately 3.     

Transmission of Haemogregarina balli from painted turtles to snapping turtles through the leech Placobdella ornata.

Six leeches (Placobdella ornata) were allowed to feed on a painted turtle (Chrysemys picta marginata) infected with Haemogregarina balli and subjected to a period of diapause before being allowed to feed on 2 laboratory-reared snapping turtles (Chelydra serpentina). Weekly examination of thin blood films revealed infections of the turtles at 130 days postfeeding. These observations provide support for broad host specificity of hemogregarine parasites of chelonians.     

First fossil gravid turtle provides insight into the evolution of reproductive traits in turtles

Here we report on the first discovery of shelled eggs inside the body cavity of a fossil turtle and on an isolated egg clutch, both referable to the Cretaceous turtle Adocus. These discoveries provide a unique opportunity to gain insight into the reproductive traits of an extinct turtle and to understand the evolution of such traits among living turtles. The gravid adult and egg clutch indicate that Adocus laid large clutches of rigid-shelled spherical eggs and established their nests near rivers, traits that are shared by its closest living relatives, the soft-shelled turtles. Adocus eggshell, however, was probably more rigid than that of living turtles, based on its great thickness and structure, features that may represent unique adaptations to intense predation or to arid nest environments. In light of the reproductive traits observed in Adocus, the distribution of reproductive traits among turtles reveals that large clutches of rigid-shelled eggs are primitive for hidden-necked turtles (cryptodirans) and that spherical eggs may have evolved independently within this group.     

Hazards Associated with the Consumption of Sea Turtle Meat and Eggs: A Review for Health Care Workers and the General Public

Sea turtle products (e.g., meat, adipose tissue, organs, blood, eggs) are common food items for many communities worldwide, despite national regulations in some countries prohibiting such consumption. However, there may be hazards associated with this consumption due to the presence of bacteria, parasites, biotoxins, and environmental contaminants. Reported health effects of consuming sea turtles infected with zoonotic pathogens include diarrhea, vomiting, and extreme dehydration, which occasionally have resulted in hospitalization and death. Levels of heavy metals and organochlorine compounds measured in sea turtle edible tissues exceed international food safety standards and could result in toxic effects including neurotoxicity, kidney disease, liver cancer, and developmental effects in fetuses and children. The health data presented in this review provide information to health care providers and the public concerning the potential hazards associated with sea turtle consumption. Based on past mortality statistics from turtle poisonings, nursing mothers and children should be particularly discouraged from consuming all sea turtle products. We recommend that individuals choose seafood items lower in the food chain that may have a lower contaminant load. Dissemination of this information via a public health campaign may simultaneously improve public health and enhance sea turtle conservation by reducing human consumption of these threatened and endangered species.     

Myxidium mackiei (Myxosporea) in Indo-Gangetic flap-shelled turtles Lissemys punctata andersonii: parasite-host interaction and ultrastructure

Myxosporeans are common parasites of fish, and uncommon parasites of amphibians, reptiles and invertebrates, that can cause significant morbidity and mortality. The common genus Myxidium infects the excretory system of turtles, yet knowledge of its pathogenicity in these hosts is limited. We offer new knowledge of morphological and ultrastructural aspects of host-parasite interactions in Myxidium infections from our recent diagnostic investigations on captive freshwater turtles listed in CITES (Appendix II). We investigated the cause of death of 2 adult Indo-Gangetic flap-shelled turtles Lissemys punctata andersonii from a zoo collection. After post-mortem examination, tissues were processed for histopathology, and special stains were used to demonstrate morphology of myxosporean spores. Additional kidney tissue, immersion-fixed in formalin, was processed for transmission electron microscopy. Both turtles were infected with a myxosporidian, Myxidium mackiei, in the kidney, which occluded 5 to 10% of the renal proximal convoluted tubules. The polysporic plasmodia contained pairs of developing and mature spores. Each mature, spindle-shaped spore had 2 asymmetric valves (1 overlapping, 1 overlapped), with 10 to 13 and 10 to 14 longitudinal ridges per valve, and 2 polar capsules each containing a polar filament with 4 to 5 turns. A pair of spores, each surrounded by a membrane-bound electron-lucent matrix, lay in an enclosing cell within the plasmodium. Regions of the parasite-host interface consisted of undulations of the parasite surface, with intense pinocytotic activity beneath, intermingled with the hosts' microvilli, and endocytotic channels at the apex of renal epithelial cells. The microvilli of the renal epithelial cells of infected tubules were frequently sheared or compressed, or occasionally missing; we did not detect other pathology induced by the parasite. Our report of M. mackiei in L. punctata is a new host record. Both individuals also had disseminated pale yellow nodules (bacterial granulomas) present in lung, heart, kidney, and skeletal muscle, and both were infected with coccidia (tentatively identified as Eimeria sp.) in multiple organs. The cause of death for one turtle was septicemia, but remained unknown for the other individual.     

A Paedomorphic parasite associated with a neotenic amphibian host: phylogenetic evidence suggests a revised systematic position for Sphyranuridae within anuran and turtle Polystomatoineans

The phylogenetic relationships of the families Polystomatidae and Sphyranuridae (subclass Polystomatoinea) within tetrapod monogenean parasites were investigated using partial 18S rDNA sequences. About 600 nucleotides of 11 species were sequenced, including 7 species of the most common subfamilies of Polystomatidae found in anurans and turtles, 1 species of the family Sphyranuridae parasitizing exclusively urodelans, and 3 species of the subclass Oligonchoinea infesting teleostean fishes. The phylogenetic analyses were performed using three reconstruction methods: neighbor-joining, maximum-parsimony, and maximum-likelihood. Polystomatoineans but not polystomatids were shown to be monophyletic. Within the polystomatoineans there are two clades: one includes the amphibian monogeneans (anuran polystomatids and urodelan sphyranurids) and the other includes the turtle polystomatids. Polystomatoineans may have coevolved with amphibian hosts, and an ancestral "polystome" dispersed at least 200 million years ago, either from the basal stem of lissamphibians or from an anuran ancestral stock, to freshwater turtles. Furthermore, the urodelan genus Sphyranura, initially assigned to the family Sphyranuridae on the basis of morphological and ontogenetic evidence, is clearly nested within polystomatids, suggesting that its systematic status must be revised. This supports recent findings which argue that species of the family Sphyranuridae may be paedomorphic parasites exclusively infesting neotenic mudpuppies.     

To Eat or not to Eat an Endangered Species: Views of Local Residents and Physicians on the Safety of Sea Turtle Consumption in Northwestern Mexico

Sea turtles have historically been an important food resource for many coastal inhabitants of Mexico. Today, the consumption of sea turtle meat and eggs continues in northwestern Mexico despite well-documented legal protection and market conditions providing easier access to other more reliable protein sources. Although there is growing evidence that consuming sea turtles may be harmful to human health due to biotoxins, environmental contaminants, viruses, parasites, and bacteria, many at-risk individuals, trusted information sources, and risk communicators may be unaware of this information. Therefore, we interviewed 134 residents and 37 physicians in a region with high rates of sea turtle consumption to: (1) examine their knowledge and perceptions concerning these risks, as a function of sex, age, occupation, education and location; (2) document the occurrence of illness resulting from consumption; and (3) identify information needs for effective risk communication. We found that 32% of physicians reported having treated patients who were sickened from sea turtle consumption. Although physicians believed sea turtles were an unhealthy food source, they were largely unaware of specific health hazards found in regional sea turtles, regardless of location. By contrast, residents believed that sea turtles were a healthy food source, regardless of sex, age, occupation, and education, and they were largely unaware of specific health hazards found in regional sea turtles, regardless of age, occupation, and education. Although most residents indicated that they would cease consumption if their physician told them it was unhealthy, women were significantly more likely to do so than men. These results suggest that residents may lack the necessary knowledge to make informed dietary decisions and physicians do not have enough accurate information to effectively communicate risks with their patients.     

Helminth component community of the loggerhead sea turtle, Caretta caretta, from Madeira Archipelago, Portugal

The helminth fauna of pelagic-stage loggerhead sea turtles, Caretta caretta, is still poorly known. Here, we describe the helminth-component community of healthy, free-ranging juvenile loggerhead sea turtles captured in the waters around Madeira Island, Portugal. Fifty-seven were used in this study. The esophagus, stomach, intestine, liver, gallbladder, spleen, kidneys, trachea, bronchi, urinary bladder, heart, left and right aortas, and coelomic cavity were macroscopically inspected; organs and tissues were removed and washed through a sieve. A search for parasites was made using a stereoscopic microscope; recovered parasites were fixed and stored in 70% alcohol until staining and identification. Prevalence, mean intensity, and mean abundance values were recorded. In total, 156 parasite specimens belonging to 9 species were found: nematodes included Anisakis simplex s.l. (larvae) and an unidentified species; digenetic trematodes present were Enodiotrema megachondrus, Rhytidodes gelatinosus, Pyelosomum renicapite, and Calycodes anthos; acanthocephalans included Bolbosoma vasculosum and Rhadinorhynchus pristis; a single cestode, Nybelinia sp., was present. Parasite infections were found to have both low prevalences and intensities. Possible reasons for this include the oligotrophic conditions of the pelagic habitat around Madeira; a 'dilution effect' because of the vastness of the area; and the small size, and thus ingestion rate, of the turtles. Results are discussed in terms of the various turtle populations that may use the waters surrounding Madeira. This work provides valuable information on the parasite fauna of a poorly known stage in the life of loggerhead sea turtles, thereby filling a fundamental gap with regard to features of the parasite fauna in this species.     

Prevalence and pathology of the nematode Heterakis gallinarum, the trematode Paratanaisia bragai, and the protozoan Histomonas meleagridis in the turkey, Meleagris gallopavo

The prevalence of infection and associated pathology induced by two helminth and one protozoan species infecting Brazilian turkeys are reported. The intestinal nematode Heterakis gallinarum appeared with a prevalence of 70% in the infected birds, without gross lesions when not associated to the protozoan Histomonas meleagridis. Histological findings in the ceca were represented by the presence of H. gallinarum worms, intense chronic diffuse inflammatory processes with mononuclear and polymorphonuclear (heterophils) leucocyte infiltrations. The prevalence of the protozoan H. meleagridis associated to H. gallinarum was of 2.5% and microscopic examination revealed a severe inflammatory process in the liver and cecum with the presence of small clear areas with round eosinophilic parasites. Gross lesions were absent in turkeys infected with the renal digenetic trematode Paratanaisia bragai; the parasite was prevalent in 20% of the cases and cross-sections of the kidneys showed a remarkable distension of the collecting ducts with several worms in the lumen. The walls of the ducts presented a discrete heterophilic infiltrate among mononuclear cells.