Invertebrates Flashcards
What happened in a study of 2PE and MS222 in HSC?
Parameters that decreased with both protocols?
2-PHENOXYETHANOL (2-PE) AND TRICAINE METHANESULFONATE (MS-222) IMMERSION ANESTHESIA OF AMERICAN HORSESHOE CRABS (LIMULUS POLYPHEMUS).
Archibald KE, Scott GN, Bailey KM, Harms CA.
J Zoo Wildl Med. 2019 Apr;50(1):96-106
Horseshoe crabs are osmoconfomers (osmolality matches surroundings)
Horseshoe crabs respond to hypoxia with a decreased blood pressure, heart rate, and gilling rate
Hemocyanin is the extracellular oxygen-carrying molecule and holds only one oxygen
The mechanism of 2-PE is unknown
Objective: Compare the effects of two immersion anesthetics, tricaine methanesulfonate (MS-222) at 1 g/L (buffered with sodium carbonate) and 2-phenoxyethanol (2-PE) at 2 mL/L, on horseshoe crabs.
Surgical anesthesia produced in 9/10 2-PE crabs and 7/10 MS-222 crabs
Parameters that decreased with both protocols: heart contractility, gilling rate, pCO2, blood pressure
Parameters that did not change with both protocols: heart rate, pH, pO2
Conclusions: Immersion in 2-PE or MS-222 produced anesthesia in American HSCs with clinically useful induction and recovery times.
Define semelparous as it pertains to GPO
What was the most common lesion in a retrospective of GPO mortality?
What parasite was found to cause branchitis? What is the treatment?
What common endosymbiotic mesozoan parasite was found in the renal sacs without inflammation?
RETROSPECTIVE REVIEW OF MORTALITY IN GIANT PACIFIC OCTOPUS (ENTEROCTOPUS DOFLEINI)
Kathryn E. Seeley, Leigh A. Clayton, Catherine A. Hadfield, Dillon Muth, Joseph L. Mankowski, Kathleen M. Kelly
J. of Zoo and Wildlife Medicine, 47(1):271-274 (2016)
Semelparous: one repro event before death- grow rapidly, die immediately after spawning
No acquired immune system, innate gets overwhelmed by poor water quality, increased bacterial loads, environ stressors
Most common lesion = ulcerations and erosions (most common on mantle or distal arms)
Two had coelomic fluid with spiral bacteria, peri-renal fluid, mineralization
Most common histologic lesions = inflammatory
Most GPOs with respiratory changes had branchitis and Ichthyobodo
Recommend monitoring siphoning effort and treat Ichthyobodo with metronidazole
Ocular lesions and renal dicyemids common
Renal dicyemids = endosymbiotic mesozoan parasite in renal sacs of many cephalopods
No inflammation associated w/ them
Summary: Branchitis was histologically evident in 14 octopuses, often in conjunction with (Ichthyobodo) or amoeba. Senescence, parasitism, and husbandry were all important contributors to mortalities.
How can death be defined for moon jellyfish following euthanasia?
Which salt is recommended for euthanasia of moon jellyfish for use in NMR?
What was observed with the other two salts in this study?
EVALUATION OF EUTHANASIA OF MOON JELLYFISH (AURELIA AURITA) USING SIMPLE SALT SOLUTIONS
Mary Doerr, Michael K. Stoskopf
J. of Zoo and Wildlife Medicine, 50(1):123-126 (2019)
Background:
- 3 neuro structures:
- Rhopalia: bell margin = pacemaker of bell contraction
- Motor nerve net = responsible for bell contraction
- Diffuse nerve net = relays signal to bell margin and rhopalia
- Ethanol and MS222 obscure results of NMR-spectroscopy-based metabolomics
- MgCl2 – used to immobilize echnioderms, cephalopods, coelenterates
- KCl – euthanasia of American lobsters
- MgSO4 – euthanize bivalve mollusks, gastropod mollusks, annelids
Key Points:
- MgCl2 at highest dose euthanized within 32 seconds without signs of distress
- KCl not suitable b/c nematocyst discharge and radial convulsing at lower concentrations + loss of tissue integrity during dissection
- MgSO4 failed to provide euthanasia
Conclusions: MgCl2 provides effective, rapid euthanasia in jellyfish
What type of weighted MRI image was best for visualizing internal organs of HSC?
Which organs were still not very well visualized?
MORPHOLOGICAL ASSESSMENT OF HORSESHOE CRABS (TACHYPLEUS TRIDENTATUS) BY USING MAGNETIC RESONANCE IMAGING
Kenji Kutara, Yumi Une, Yoshinobu Fujita
J. of Zoo and Wildlife Medicine, 50(3):742-748 (2019)
• T2 weighted gave best images (best tissue contrast)
• Muscles able to be distinguished but not muscles of legs (resolution too low)
• Liver and CV system not distinguished because isointense
Take Home Message: Use T2 weighted MRI to visualize internal organs of horseshoe crabs; does not do a good job at evaluating cardiovascular or liver
How is anesthetic induction defined in sea urchins?
What immersion anesthetic has been used to safely sedate and anesthetize sea urchins?
TRICAINE METHANESULFONATE (MS-222) SEDATION AND ANESTHESIA IN THE PURPLE-SPINED SEA URCHIN (ARBACIA PUNCTULATA)
Applegate Jr JR, Dombrowski DS, Christian LS, Bayer MP, Harms CA, Lewbart GA.
Journal of Zoo and Wildlife Medicine. 2016 Dec;47(4):1025-33.
Effective anesthesia in other aquatic invertebrates:
• Isoflurane in octopus and tarantulas
• Alfaxalone in blue crabs
• KCl and MgCl in other inverts
Sea urchin Anesthesia:
• Sedation: loss of righting reflex with retained tactile spine response
• Induction: loss of righting reflex, release from a vertical surface
• Anesthesia: loss of righting reflex and loss of tactile spine response.
• Recovery: voluntary return to oral recumbency
• MS-222 @ 0.4 g/L → sedation
• MS-222 @ 0.8 g/L → anesthesia
• More unplanned spawning than at lower dose
• Recovery time was not dose dependent
• Induction with MS-222 was faster than with KCl in other studies
• No mortalities
Describe the technique for intracoelomic injection/hemolymph collection in sea urchins.
What was observed in a PK study of enrofloxacin and metabolite ciprofloxacin in urchins by injection and immersion?
POPULATION PHARMACOKINETICS OF ENROFLOXACIN AND ITS METABOLITE CIPROFLOXACIN IN THE GREEN SEA URCHIN (STRONGYLOCENTROTUS DROEBACHIENSIS) FOLLOWING INTRACOELOMIC AND IMMERSION ADMINISTRATION
Phillips BE, Harms CA, Lewbart GA, Lahner LL, Haulena M, Rosenberg JF, Papich MG.
Journal of Zoo and Wildlife Medicine. 2016 Mar;47(1):175-86.
Intracoelomic administration / hemolymph collection: insert needle perpendicular through peristomal membrane lateral to mouth at edge of the test (exoskeleton)
• No adverse effects
• Measurement of ciprofloxacin metabolite was low and inconsistent
• Immersion Cmax occurred >9 hr after completion of treatment
• Both treatments exceeded MIC for Vibrio and Pseudomonas and other pathogenic bacteria for days
Conclusions: Treatment of 10 mg/kg intraceolomic every 5 days or immersion 10 mg/L every 3 days for 6 hr may be effective treatment for urchins.
Describe technique for collection of water vascular system fluid vs introcoelomic injection in sea stars.
What was observed in a PK study of enrofloxacin with intracoelomic vs immersion administration in sea stars?
Population pharmacokinetics of enrofloxacin in purple sea stars (Pisaster ochraceus) following an intracoelomic injection or extended immersion
Justin F. Rosenberg DVM; Martin Haulena DVM, MSc; Brianne E. Phillips DVM; Craig A. Harms DVM, PhD; Gregory A. Lewbart MS, VMD; Lesanna L. Lahner DVM, MPH; Mark G. Papich DVM, MS
American Journal of Veterinary Research November 2016, Vol. 77, No. 11, Pages 1266-1275
Background:
- Sea star = echinoderm
- Coelomic fluid, water vascular fluid system, and hemal fluid are 3 separate compartments
- Collection of water vascular system fluid: aboral aspect of limbs 1cm from distal tip, 27g needle/1ml syringe
- Intracoelomic injections – 27g needle on aboral surface at base of limb near junction w/ central disc
Key Points:
• Intraceolomic was higher levels (Cmax) and longer half life than immersion
• No adverse effects
• Enrofloxacin present for > 126 hours (stopped testing)
• Ciprofloxacin barely detectable after injection
• Not detectable in green urchins
• Other inverts can convert enro to cipro: Chinese mitten-handed crabs, giant freshwater prawns, ridgetail white prawns, Chinese shrimp
• No adverse effects
Conclusions: Intracoelomic administration of enrofloxacin in purple sea stars had higher levels than immersion.
What is the causative agent of aspergillosis in sea fans in the FL keys?
Aspergillus sydowii
What is the oxygen binding protein in most aquatic inverts? Which group generally lacks an O2 binding pigment?
Hemocyanin
Bivalves - Majority lack oxygen binding pigment, O2 dissolved directly into hemolymph; Except the Marine Ark Anadara trapezia (multiple hemoglobins)
Causative agent of Marteiliosis in bivalves?
Is this a reportable dz?
Marteiliosis (protist, paramyxida) aka Aber dz aka Aq dz aka digestive gland dz
Marteilia refringens (Eu, Americas)
Marteilia sydneyi (Aus)
Enters host via epithelium of palps and gills
Protozoa develop and proliferate in GI
IM host lifecycle - Probably copepods
Death generally May-Aug of second growth year
Causative agent of Bonamiosis in oysters?
Is this a reportable dz?
Bonamiosis (haplosporidian) AKA microcell disease, hemocyte disease, winter mortality
Disease of oysters – Flat oyster, Foveax Strait oyster, Ostrea denselammellosa
Associated with haplosporidians:
Bonamia ostreae
Bonamia exitiosus
Mikrocytos roughleyi
Very wide geographic range: NA, Europe, Australia
Profound economic losses
Mortalities up to 80%
Dual infection with other protozoa enhance mortality
Infection intensity greatest in females after spawning
Reportable by the OIE
CS: Discoloration of digestive gland, mantle, gills, decline in body condition
Dx via hemolymph smears, tissue imprints, histology
Bonamia organisms 2-5 microns in size
Heart tissue imprints
Massive tissue hemocyte infiltration
PCR, in situ hybridization for monoclonal antibodies
Combined use of heart tissue smears and PCR enhances sensitivity
Infection is contagious
Early harvesting at 15-18 months of production and subtidal culture may minimize impact
Breeding programs trying to produce Bonamia ostrea resistant oyster stocks
What are two reportable diseases of bivalves?
Marteilliosis (paramyxida), Bonamiosis (haplosporidian); Both protozoal dz
What compound is extracted from HSC for endotoxin detection in human medicine?
Limulus amebocyte lysate
What are the three body sections of HSC?
How are males distinguished from females?
Cephalothorax (prosoma), abdomen (opisthosoma), telson (tail)
Males have a single pair of modified chelicera - large bulbous claspers to grasp female
PK of enrofloxacin and ciprofloxacin in HSC?
Single injection 5 mg/kg in dorsal cardiac sinus lasted 120 hrs, no adverse effects, MIC reached.
A recent study described hemolymph collection in emperor scorpions.
How were they restrained?
- How effective was it?
- How long did recovery take?
- How did this differ from other scorpion species?
Describe hemolymph collection in this species
- What was the colleciton site?
- How much hemolymph was collected?
- What is hemolymph?
- Describe its role in the scoprion circulatory system.
JZWM 2022 53(3) 573-577
HEMOLYMPH COLLECTION AND ISOFLURANE ANESTHESIA OF THE EMPEROR SCORPION (PANDINUS IMPERATOR)
Abstract: With captive invertebrates increasing in popularity, a safe and effective anesthesia protocol is required for their veterinary care. This study investigated the safety and efficacy of 5% isoflurane anesthesia and 0.26 ml hemolymph collection in subadult to adult emperor scorpions (Pandinus imperator; n = 14). Each scorpion was placed in a 2-L anesthetic chamber and induced with 5% isoflurane at a flow rate of 2 L/min oxygen. Anesthetic depth was assessed by rolling the animal into dorsal recumbency and rating the response from one to four: 1 = successful coordinated righting movement, 2 = successful uncoordinated righting movement, 3 = unsuccessful righting movement, 4 = no movement. Ratings 3 and 4 were both adequate for the hemolymph collection. The mean induction time was 4.2 min to reach a level 3 depth and 10.5 min to reach a level 4 depth. All animals were clinically normal for at least 1 yr post experiment. A mean hemolymph volume drawn of 1.92% of body weight, and an anesthetic induction with 5% isoflurane, appear safe in subadult to adult emperor scorpions.
Intro
- Background anatomy info:
– Scorpions are segmentally divided into the prosoma, mesosoma, and metasoma.
– The semiclosed circulatory system is composed of a dorsosagitally located heart in the mesosoma.
– The contractions of the heart, in conjunction with body and appendage movement, circulates the hemolymph and its constituents (e.g., hemocytes, nutrients, dissolved gasses)
- Hemolymph interacts with the respiratory system for gas exchange at four pairs of book lungs located on the ventral surface of tergites 10–13 in the emperor scorpion.
- Each book lung is an invagination of the exoskeleton, creating a cave-like antrum with an external opening called the spiracle. Within each antrum are thin parallel sheets of tissue lamellae for gas exchange with the hemolymph
- The objective of this study was to determine if anesthesia with 5% isoflurane, followed by 0.26 ml hemolymph collection, is a safe and effective protocol for subadult to adult emperor scorpions.
M&M
- n=14 subadult to adult emperor scorpions
- The scorpion was placed in a 2L anesthesia chamber on 5% isoflurane (O2 flow 2L/min)
- The scorpions were challenged at 30-s intervals by rotating the chamber 908 and attempting to roll the animal into dorsal recumbency.
- At each interval, the animal’s response to stimuli was rated on a scale of one to four: 1 = successful coordinated righting movement, 2 = successful uncoordinated righting movement, 3 = unsuccessful righting movement, 4 = no movement.
- The anesthetic gas was turned off after the animal had three consecutive time points at a rating of 4 or at 30 min, whichever came first
- A pericardial hemolymph sample was collected: at the dorsal midline, between mesosoma (abdomen) tergites five and six, a volume of 0.26 ml was collected.
- Each scorpion was placed in a plastic container with a screened top for room-air recovery.
- During recovery, all scorpions were challenged every 30 s and rated on the same response scale as used for anesthetic induction.
- The scorpions were considered recovered after three consecutive ratings of 1
Results
- The average time to a rating of 3 was 4.2 min and rating 4 being 10.5 min
- The five scorpions that did not receive a rating of 4 were nonetheless considered at an anesthetic plane safe enough for hemolymph collection after 30 min
- The average time to recovery for all scorpions, following discontinuation of anesthetic gas, was 12.7 min
- The average hemolymph drawn as a percent of body weight was 1.92%
- Hemolymph collection was possible in all scorpions, and no obvious signs of distress were noted during induction or recovery
- All scorpions alive 1 year post study
Discussion
- This study considered a rating of 3 or 4 safe for minor procedures such as hemolymph collection
- A previous study showed 5% isoflurane anesthesia in Asian forest scorpions was effective, however during sedation a dangerous hyperexcitatory phase was observed
- This was contrasted by the dictator scorpions (Pandinus dictator) in the same study having a smooth induction, similar to the emperor scorpion in the current study
- This may mean that scorpions in the same genus, Pandinus, have similar induction reactions to 5% isoflurane
- Importantly, differences in average induction time and recovery may suggest extrapolation between genera or species within Scorpiones is of limited value.
Takeaway: This study shows that hemolymph collection up to 1.92% of body weight and anesthesia with 5% isoflurane up to 30 min does not noticeably impact behavior or survival in subadult to adult emperor scorpions
