A practical guide to necropsy of the elasmobranch chondrocranium and causes of mortality in wild and aquarium-housed California elasmobranchs.

Elasmobranchs are common, iconic species in public aquaria; their wild counterparts are key members of marine ecosystems. Post-mortem examination is a critical tool for disease monitoring of wild elasmobranchs and for management of those under human care. Careful necropsy of the head, with a focus on clinically relevant anatomy, can ensure that proper samples are collected, increasing the chance of presumptive diagnoses prior to slower diagnostic workup. Immediate feedback from a thorough head necropsy allows for faster management decisions, often identifying pathogens, routes of pathogen entry, and pathogenesis, which are current shortcomings in published literature. This article proposes a protocol for necropsy of the elasmobranch chondrocranium, emphasizing unique anatomy and careful dissection, evaluation, and sampling of the endolymphatic pores and ducts, inner ears, brain, and olfactory system as part of a complete, whole-body necropsy. Extensive use of cytology and microbiology, along with thorough sample collection for histology and molecular biology, has proven effective in identifying a wide range of pathogens and assisting with characterization of pathogenesis. The cause of mortality is often identified from a head necropsy alone, but does not replace a thorough whole-body dissection. This protocol for necropsy and ancillary diagnostic sample collection and evaluation was developed and implemented in the necropsy of 189 wild and aquarium-housed elasmobranchs across 18 species over 13 years (2011-2023) in California. Using this chondrocranial approach, meningoencephalitis was determined to be the primary cause of mortality in 70% (118/168) of stranded wild and aquarium-housed elasmobranchs. Etiology was largely bacterial or protozoal. Carnobacterium maltaromaticum bacterial meningoencephalitis occurred in salmon sharks (Lamna ditropis), shortfin mako sharks (Isurus oxyrinchus), common thresher sharks (Alopias vulpinus), and one Pacific electric ray (Tetronarce californica). Miamiensis avidus was the most common cause of protozoal meningoencephalitis and found almost exclusively in leopard sharks (Triakis semifasciata) and bat rays (Myliobatis californica) that stranded in San Francisco Bay. Bacterial pathogens were found to use an endolymphatic route of entry, while protozoa entered via the nares and olfactory lamellae. Trauma was the second most common cause of mortality and responsible for 14% (24/168) of wild shark strandings and deaths of aquarium-housed animals.

spaB-positive Erysipelothrix rhusiopathiae, a novel teleost pathogen isolated from cultured barramundi.

Members of the genus Erysipelothrix are emergent pathogens of cultured eels, as well as several characid and cyprinid species. Since 2013, E. rhusiopathiae has been reported from diseased barramundi (Lates calcarifer) cultured in North America; we recovered 8 E. rhusiopathiae isolates from diseased fish during different outbreaks from the same farm. The E. rhusiopathiae isolates from barramundi were compared phenotypically and genetically to E. piscisicarius isolates characterized from ornamental fish and E. rhusiopathiae recovered from aquatic and terrestrial animals. All barramundi isolates were PCR-positive for the surface protective antigen type B (spaB) gene, and shared ≥ 99.7% sequence similarity among concatenated multilocus sequence analysis gene sequences, indicating a high degree of genetic homogeneity. These isolates were > 99% similar to other spaB-positive isolates from marine invertebrates and marine mammals, consistent with findings for other spa types. The spaA and spaB isolates shared < 98% similarity, as well as < 90% similarity with spaC-positive E. piscisicarius. Similar clonality among the spaB isolates was observed using repetitive element palindromic PCR. In experimental intracoelomic injection challenges conducted to fulfill Koch postulates, 67% of exposed tiger barbs (Puntigrus tetrazona) died within 14 d of challenge. Our study supports previous work citing the genetic variability of Erysipelothrix spp. spa types and the emergence of members of the genus Erysipelothrix as nascent fish pathogens.

DEXMEDETOMIDINE AND MIDAZOLAM INTRAMUSCULAR SEDATION IN BROWNBANDED BAMBOO SHARKS (CHILOSCYLLIUM PUNCTATUM).

Bamboo sharks are some of the most common elasmobranch species in zoos and aquaria and are frequently sedated for medical exams, treatments, and research. This study investigated the use of an IM sedation protocol of a single dose of dexmedetomidine (0.05 mg/kg) and midazolam (2.0 mg/kg) in brownbanded bamboo sharks (Chiloscyllium punctatum). Sharks were serially monitored every 5 min for heart rate, branchial beats, righting reflex, coelomic response, cloacal response, pelvic fin reflex, response to noxious stimulus, voluntary movement, and ability to swim. This sedation dose was effective at rapidly and significantly decreasing responses to tactile and noxious stimuli with minimal respiratory depression and was quickly reversible with atipamezole (0.5 mg/kg) and flumazenil (0.05 mg/kg). Sedated sharks developed a mild metabolic acidosis evidenced by a significant increase in lactic acid (mean < 0.37 mmol/L presedation, 4.2 mmol/L after reversal) and decrease in blood pH (mean 7.464 presedation, 7.277 after reversal); however, clinical intervention was not required. This protocol should be further investigated in different elasmobranch species but is promising for providing sedation for noninvasive procedures in brownbanded bamboo sharks.

Pathology, microbiology, and genetic diversity associated with Erysipelothrix rhusiopathiae and novel Erysipelothrix spp. infections in southern sea otters (Enhydra lutris nereis).

Erysipelothrix spp., including E. rhusiopathiae, are zoonotic bacterial pathogens that can cause morbidity and mortality in mammals, fish, reptiles, birds, and humans. The southern sea otter (SSO; Enhydra lutris nereis) is a federally-listed threatened species for which infectious disease is a major cause of mortality. We estimated the frequency of detection of these opportunistic pathogens in dead SSOs, described pathology associated with Erysipelothrix infections in SSOs, characterized the genetic diversity and antimicrobial susceptibility of SSO isolates, and evaluated the virulence of two novel Erysipelothrix isolates from SSOs using an in vivo fish model. From 1998 to 2021 Erysipelothrix spp. were isolated from six of >500 necropsied SSOs. Erysipelothrix spp. were isolated in pure culture from three cases, while the other three were mixed cultures. Bacterial septicemia was a primary or contributing cause of death in five of the six cases. Other pathology observed included suppurative lymphadenopathy, fibrinosuppurative arteritis with thrombosis and infarction, bilateral uveitis and endophthalmitis, hypopyon, petechia and ecchymoses, mucosal infarction, and suppurative meningoencephalitis and ventriculitis. Short to long slender Gram-positive or Gram-variable bacterial rods were identified within lesions, alone or with other opportunistic bacteria. All six SSO isolates had the spaA genotype-four isolates clustered with spaA E. rhusiopathiae strains from various terrestrial and marine animal hosts. Two isolates did not cluster with any known Erysipelothrix spp.; whole genome sequencing revealed a novel Erysipelothrix species and a novel E. rhusiopathiae subspecies. We propose the names Erysipelothrix enhydrae sp. nov. and Erysipelothrix rhusiopathiae ohloneorum ssp. nov. respectively. The type strains are E. enhydrae UCD-4322-04 and E. rhusiopathiae ohloneorum UCD-4724-06, respectively. Experimental injection of tiger barbs (Puntigrus tetrazona) resulted in infection and mortality from the two novel Erysipelothrix spp. Antimicrobial susceptibility testing of Erysipelothrix isolates from SSOs shows similar susceptibility profiles to isolates from other terrestrial and aquatic animals. This is the first description of the pathology, microbial characteristics, and genetic diversity of Erysipelothrix isolates recovered from diseased SSOs. Methods presented here can facilitate case recognition, aid characterization of Erysipelothrix isolates, and illustrate assessment of virulence using fish models.

Limb girdle muscular dystrophy type 2A presenting with cardiac arrest.

The occurence of respiratory failure in progressive neuromuscular disorders is well recognized. This failure is observed most commonly in Duchenne dystrophy but sometimes occurs in Becker's, limb-girdle, and facioscapulohumeral dystrophies. Patients usually present acutely or subacutely with cyanosis and cor pulmonale, with severe decompensation often being precipitated by an acute intercurrent infection. However, cardiopulmonary arrest is an uncommon presentation. A male diagnosed with limb-girdle muscular dystrophy type 2A who presented with cardiopulmonary arrest that was precipitated by an upper respiratory tract infection is presented. The nocturnal application of noninvasive intermittent positive pressure ventilation with a bilevel positive airway pressure (Bi-PAP) device improved his symptoms and quality of life without resorting to more-invasive and more-restrictive forms of support. This report demonstrates an unusual presentation of limb-girdle muscular dystrophy and documents that nocturnal nasal administration of continuous airway pressure using the Bi-PAP device may be sufficient to maintain adequate ventilation in such patients.

The clearance of proteoglycan-associated human recombinant tissue factor pathway inhibitor (h-rTFPI) in rabbits: a complex formation of h-rTFPI with factor Xa promotes a clearance rate of h-rTFPI.

The very rapid clearance of human recombinant tissue factor pathway inhibitor (h-rTFPI) may result from its binding to vascular proteogly can and LDL receptor-related protein (LRP). To investigate the effect of factor Xa on the clearance of h-rTFPI, we developed a specific ELISA for h-rTFPI/factor Xa complex, and compared the pharmacokinetic parameters of h-rTFPI/factor Xa complex and the clearance rate of the cellular proteogly can-associated h-rTFPI/factor Xa complex with those of h-rTFPI by itself in rabbits. We found that the h-rTFPI/factor Xa complex disappeared from circulation at a rapid rate of clearance, having pharmacokinetic parameters similar to those of non-complexed h-rTFPI. After the rapid disappearance of the h-rTFPI complex from plasma, an intravenous injection of heparin resulted in a release of h-rTFPI/factor Xa complex into plasma. However, the recovery of heparin-releasable h-rTFPI/factor Xa decreased significantly in a time-dependent manner. Therefore, we examined the half-life of proteogly can-associated h-rTFPI/factor Xa and determined it to be 51 min, which was significantly shorter than that of h-rTFPI by itself (107 min). These results suggest that a complex formation of h-rTFPI with factor Xa promotes a clearance of proteogly can-associated h-rTFPI existing in the liver and kidney.

[Brain CT and MRI findings in fat embolism syndrome].

To elucidate brain CT and MRI findings in fat embolism syndrome (FES), we retrospectively analyzed images from 5 patients with FES during the acute and subacute stages. Brain CT examinations demonstrated brain edema in 2 patients and transient spotty low density lesions in 2 patients. Three patients showed no abnormalities. Brain MRI, however, showed brain abnormalities in all patients during the acute stages. These were revealed as spotty high signal intensity lesions on T2WI, and some showed low intensity on T1WI. These spotty lesions were considered to reflect edematous fluid occurring as a result of the unique pathophysiological condition of FES. While the spotty high signal intensity lesions on T2WI were distributed in the cerebrum, cerebellum, brain stem, thalamus, basal ganglia, internal capsule and corpus callosum, cerebral and cerebellar spotty lesions were characteristically located along the boundary zones of the major vascular territories. This characteristic location might be induced by a hypoxic brain condition in FES because the numerous fat globules present in this condition can block entire brain capillaries. This characteristic signal location on T2WI is a useful indicator for differentiating FES from the primary intra-axial brain injury in patients with multifocal trauma.

Protective effect of dipyridamole against lethality and lipid peroxidation in liver and spleen of the ddY mouse after whole-body irradiation.

The effects of dipyridamole on radiation damage in the mouse were investigated. Dipyridamole (i.p. 2 mg/mouse) administered 1 h before exposure, protected against gamma-irradiation. Pretreatment significantly decreased the death rate at 30 days from 89 to 33% (p<0.001) after 9 Gy whole-body irradiation. LD50 at 30 days was increased from 6.67 to 7.65 Gy in the dipyridamole pretreated group. The level of thiobarbituric acid reactive substances (TBARS) in the liver and spleen, a measure of free radical initiated liver peroxidation, increased 155, 193, 195, and 236% of control (without irradiation) in liver, and 132, 146, 168, and 276% of control (without irradiation) in spleen on days 2, 4, 7, and 10 after 9 Gy of whole-body irradiation respectively. The TBARS levels in both liver and spleen 2 days after irradiation were reduced to 73 +/- 7 and 60 +/- 19% respectively after dipyridamole treatment (2 mg/mouse, i.p. injection 1 h before exposure). In electron microscopic studies, mitochondria and endoplasmic reticulum in the irradiated mouse liver were swollen, but otherwise appeared normal after dipyridamole treatment. These results suggest that dipyridamole has a protective effect on animal survival 30 days after 60Co gamma-irradiation and inhibits lipid peroxidation - which is thought to play a part in the radiation injury in mouse liver and spleen.

Bronchioloalveolar adenoma of the lung: CT-pathologic correlation.

PURPOSE: To correlate the appearance of bronchioloalveolar adenoma (BAA) of the lung at computed tomography (CT) with its pathologic features. MATERIALS AND METHODS: Nine small pulmonary nodules with ground-glass attenuation were found at CT in 668 patients with lung carcinoma. Seven of these nine lesions were histopathologically diagnosed as BAA in four patients (three men and one woman, aged 66-77 years) (three lesions in one man, two lesions in the woman, and one lesion each in the remaining two men) and are the subject of this study. RESULTS: BAA of the lung appeared at CT as a small pulmonary nodule with ground-glass attenuation; microscopic examination revealed hyperplasia of the alveolar cuboidal cells on hyperplastic alveolar septa. The CT findings depicted partial reduction of the alveolar air spaces owing to an increase in cellular components within the lesion. Adenocarcinoma cells were also seen within the BAA lesion in one patient. CONCLUSION: In a patient with known lung carcinoma, a small nodule with ground-glass attenuation simulating a focal lesion of pulmonary interstitial disease must be investigated to rule out BAA or multicentric adenocarcinoma.

[CT findings of sinonasal and orbital Wegener's granulomatosis].

We reviewed the CT findings of four cases of Wegener's granulomatosis that presented as inflammatory masses in the sinonasal cavity or orbit. In the present study, an infiltrative nature and homogeneous texture with contrast enhancement were typical of the masses. In addition, the masses were frequently accompanied by infiltration of the pterygopalatine fossa or destruction of adjacent bone. However, no pathognomonic findings were observed.

[CT findings of pulmonary inflammatory pseudotumors (plasma cell granulomas)].

Pulmonary inflammatory pseudotumor (plasma cell granuloma) is not a true neoplastic lesion, but is composed of a variety of inflammatory cells, predominantly plasma cells. The chest X-ray features resemble those of malignant lung tumors; therefore, CT is often necessary for further evaluation. We report the CT features of five cases with histologically proved pulmonary inflammatory pseudotumor, which can be summarized as follows: a solitary round or oval parenchymal mass with regular or irregular margin, and with or without calcifications. The calcifications are useful for differential diagnosis if present, but they are usually non-specific in shape and configuration. The mean CT attenuation value of the major portion of the mass was increased from 41 HU to 78 HU by the injection of contrast material. In one case, a linear extension of the lesion was seen from the mass to the lung hilum. In three cases, satellite lung nodules were seen. One of these nodules was also proved histopathologically to be inflammatory pseudotumor. The biopsy specimens obtained by using a 20 gauge cutting-needle and an automated biopsy gun were satisfactory for histopathological diagnosis.

[CT and MR imaging of desmoid tumors--comparison of two modalities].

As desmoid tumors invade locally and postoperative recurrence is common, accurate diagnosis of the extent of the tumor is needed prior to surgery. CT and/or MRI evaluation of tumor extension was retrospectively studied in eight patients with desmoid tumors, and the results were correlated with the histopathological findings. All tumors were completely resected even in patients who were evaluated by CT alone. However, the delineation of tumor and local invasion were not clearly demonstrated by CT. On the other hand, the delineation of tumor and local invasion were well visualized on MRI. The MRI picture of desmoid tumors was mainly composed of two different areas of signal intensity. The area of hypointensity in both T 1- and T 2-weighted images was found to have abundant collagen fibers, while the area of isointensity or slight hyperintensity in T 1-weighted images and hyperintensity in T 2-weighted images was found to have fibroblasts. In conclusion, MRI is better suited to the evaluation of patients with desmoid tumors than CT.

[CT features of chronic maxillary osteomyelitis in adults].

CT findings were reviewed in four adult patients with chronic maxillary osteomyelitis (CMO) that was histologically proved. The CT features of CMO included bone destruction and soft tissue mass, predominantly in the inferior portion of the maxillary antrum (all 4 cases), thickening of the antral wall (3 cases) and abnormal soft tissue around the antrum associated with or without bony wall destruction (3 cases). CMO could not be distinguished from cancer of the maxillary antrum on CT because of the similar findings. However, abnormal soft tissue around the antrum together with an undestructed bony antral wall may be useful for differentiating the two diseases.

[CT findings of mucin-producing pancreatic cancer--differentiation from chronic pancreatitis].

Mucin-producing pancreatic cancers (MPPC), which include mucinous adenocarcinoma, papillary adenocarcinoma and cystadenocarcinoma, are radiographically characterized by diffuse or localized dilatation of the main pancreatic duct due to excessive mucin production. Therefore, MPPC are occasionally difficult to distinguish from chronic pancreatitis on CT unless the primary pancreatic lesion is visualized. We compared five cases of MPPC with five cases of chronic pancreatitis with marked duct dilatation to determine differences in CT images between the two diseases. There was no significant difference between the two diseases in the nature of duct dilatation (size, extent, contour) or parenchymal changes (atrophy, enlargement, calcification, cystic lesion). However, dilatation of the intramural duct was characteristically observed in MPPC but not in chronic pancreatitis. Papillary masses in the pancreatic duct, when observed, were another finding specific to MPPC.

[CT-guided percutaneous lung biopsy using a cutting needle and an automated biopsy gun. Comparison with lung biopsy using a manual aspiration needle].

Twenty-five percutaneous lung biopsies using a 20-gauge cutting needle and automated biopsy gun (ABG) were performed under CT guidance in 25 patients with thoracic lesions. This procedure was compared with that using a 21-gauge manual aspiration needle in 36 patients (40 examinations, 37 lesions) in terms of success rate, rate of correct diagnosis, mean examination time and rate of complications. Specimens obtained from lung biopsy were graded by a histopathologist according to quality and quantity from 0 to 4 (pathological score). There were no statistically significant differences between the two procedures in terms of success rate, rate of correct diagnosis and rate of complications; only the time required was significantly different. However, sufficient biopsy material and a mean pathological score of G-II 2.8 (that of G-I was 1.9, p less than 0.05) could be obtained by the biopsy procedure using the cutting needle. The above results indicated that aspiration needle biopsy was adequate for lung biopsy, but that a cutting needle and ABG should be used when a good biopsy specimen is needed for tissue diagnosis.

[New oral contrast material for gastrointestinal MR examination--green tea mixed with sodium alginate].

We found incidentally the stomach filled with the material of high signal intensity on T1 weighted images. The material was a mixture of green tea and sodium alginate (drug for peptic ulcer). We proved that T1 and T2 were significantly shortened by the mixture in the experimental study. This was successfully used for the patients.

[Study on insulin resistance in rats treated with estrogen and progesterone--assessment with the euglycemic glucose clamp technic].

To clarify the mechanism of insulin resistance in pregnancy, we have used the euglycemic glucose clamp technique in estradiol(E) treatment(n = 6), progesterone(P) treatment (n = 28), and Control(n = 29) female rats. E(10 micrograms/day) and P(10 mg/day) were injected subcutaneously into female rats for 14 days, to increase E and P concentrations to pregnant levels. Glucose production and glucose utilization were measured by using [3-3H]-glucose. The results were as follows, 1) Glucose production was almost suppressed at hyperinsulinemia(11,000 microU/ml) both Control and P treatment rats. Then at hyperinsulinemia, glucose utilization rate was almost equal to glucose infusion rate. 2) In P treatment rats glucose utilization was significantly lower (p less than 0.05) than in Control rats at hyperinsulinemia (11,000 microU/ml). 3) In P treatment rats glucose infusion rate was significantly lower than in Control rats at plasma insulin concentrations of 1,000 microU/ml(p less than 0.02), and 11,000 microU/ml(p less than 0.01), and lower than in E treatment rats at plasma insulin concentrations of 11,000 microU/ml(p less than 0.05). 4) In a dose-response curve for the effects of four different concentrations of insulin on glucose infusion rate, the insulin resistance induced by progesterone is characterized by a decreased responsiveness to insulin. The results suggest that progesterone may play an important role in inducing insulin resistance in pregnancy.

[Prevention of postoperative infection following cardiac catheterization in pediatric field. Study of air-borne bacteria in X-ray room and evaluation of an antibiotic used for prevention of postoperative infection].

As part of preventive measures against postoperative infection following cardiac catheterization in infants with cardiac diseases, especially falling bacteria in X-ray room was studied. Moreover, a synthetic penicillin, ticarcillin (TIPC), was used as preventive antibiotic against postoperative infections due to falling bacteria which probably contaminate the air in the X-ray examination room, and the efficacy and side effects of the drug were observed. As result, coagulase-negative Staphylococcus was detected the most, followed by Micrococcus and then by fungus. The number of these 3 organisms corresponded to 90.3% of the total number of falling bacteria detected during operation. The number of falling bacteria during operation was 5.1 times larger than that before operation. Taking into account normal flora of skin, falling bacteria present in the X-ray room and causative organisms of bacterial endocarditis, TIPC was administered to 30 cases intravenously 5 times at a dose of 30 mg/kg every 8 hours for the purpose of preventing possible postoperative infections following cardiac catheterization. The drug was effective to prevent such infections in all cases. No side effects were noted in any case, in peripheral blood and hepatic function tests and other observations.