Amphibian Leukocyte Profiles Home

The table below lists the leukocyte profiles (percentage of cells) for amphibians from published and unpublished sources. The mean percentage of each cell type (lymphocytes, neutrophils, eosinophils, basophils and monocytes) is given, along with the neutrophil/lymphocyte ratio, which is useful to gauge the stress level of the animals (see review paper in further reading page). Reference range of N/L ratios is shown below table. If the study was an experiment, only the values from control animals is listed. References are listed below the table. This list was compiled after many long hours at a library and at the microscope. If you know of other publications, or have data to add, contact akdavis(at)uga.edu.

Last updated Dec 30 2011

Species Group Age N Status Lympho. Neutro. Eosino. Baso. Mono. N/L Ratio Source Notes
Grey Tree Frog (Hyla versicolor) Anuran Adult 11 Wild 83.3 9.4 2.6 3.2 1.4 0.13 Davis, upubl. data collected from a site at Whitehall Forest near Athens GA
Cuban Tree Frog (Osteopilus septentrionalis) Anuran Adult 7 Captive 63.6 24.6 6.2 1.5 4.2 0.39 Davis & Horan, unpubl. data non-breeding
Northern Cricket Frog (Acris c. crepitans) Anuran Adult 79 Wild 68.3 22.4 1.6 5.0 2.7 0.33 Davis & Durso 2009 non-breeding, from two ponds in northeast GA
American Bullfrog (Rana catesbeiana) Anuran Adult 14 Captive 62.9 22.0 8.9 2.5 0.6 0.35 Cathers et al. 1997
American Bullfrog (Rana catesbeiana) Anuran Adult 302 Captive 26.8 60.9 5.8 3.5 2.9 2.27 Coppo et al. 2005 all sampled 24hrs after transport
American Bullfrog (Rana catesbeiana) Anuran Larvae 40 Wild 73.0 23.8 3.6 2.3 0.3 0.33 Davis 2009 all pre-metamorphic
American Bullfrog (Rana catesbeiana) Anuran Adult 7 Wild 67.4 8.8 11.6 10.5 1.8 0.13 Davis 2009 from multiple ponds in northeast GA
American Bullfrog (Rana catesbeiana) Anuran Larvae 12 Reared 91.8 0.8 1.8 5.6 0.0 0.01 Rocha et al. 2010 Farm-reared animals, all brought to lab and acclimated for 7 days
Leopard Frog (Rana pipiens) Anuran Adult 50 Captive 53.4 26.5 7.3 4.4 11.0 0.50 Rouf 1969
Leopard Frog (Rana pipiens) Anuran Adult 18 Captive 55.4 11.3 10.1 19.2 4.1 0.20 Maniero & Carey 1997
Leopard Frog (Rana pipiens) Anuran Adult 14 Captive 25.4 61.8 7.0 1.8 5.2 2.43 Bennett & Alspaugh 1964 not clear what caused these unusual values
Leopard Frog (Rana pipiens) Anuran Adult 93 Unknown 64.5 20.7 8.0 6.7 0.0 0.32 Kaplan 1952
Leopard Frog (Rana pipiens) Anuran Adult 82 Wild 36.0 38.7 18.6 5.9 0.8 1.08 Shutler and Marcogliese 2011  
Southern Leopard Frog (Rana sphenocephala) Anuran Adult 69 Reared 63.6 13.8 4.2 12.4 5.9 0.22 Davis, unpubl. data just metamorphed
Southern Leopard Frog (Rana sphenocephala) Anuran Adult 61 Reared 60.7 23.9 1.99 9.9 3.5 0.39 Davis, unpubl. data just metamorphed
Edible frog (Rana esculenta) Anuran Adult * Unknown 75.2 17.1 5.7 1.9 0.0 0.23 Friedsohn 1910, cited in Jordan 1938
Edible frog (Rana esculenta) Anuran Adult 136 Wild 57.6 15.2 14.4 12.4 0.5 0.26 Romanova &Romanova 2003
Green frog (Rana clamitans) Anuran Adult 35 Wild 66.0 16.0 17.0 1.0 1.0 0.24 Shutler et al 2009 population in NS, Canada
South American Toad (Bufo arenarum) Anuran Adult 12 Captive 60.9 27.3 3.7 3.8 1.7 0.45 Chiesa et al. 2006
South American Toad (Bufo arenarum) Anuran Adult 24 Wild 64.0 20.9 13.7 0.0 1.3 0.33 Cabagna et al. 2005
American Toad (Bufo americanus) Anuran Adult 27 Wild 20.0 68.0 3.3 7.4 1.5 3.40 Forbes et al. 2006 all toads were breeding
American Toad (Bufo americanus) Anuran Adult 50 Wild 75.9 7.0 5.9 10.5 0.6 0.09 Davis & DeVore, unpubl. data all 1 year old, non-breeding
Colorado River toad (Bufo alvarius) Anuran Adult * Wild 37.0 48.0 9.0 1.0 5.0 1.30 Cannon & Cannon 1979 captured in August
Fowlers Toad (Bufo fowleri) Anuran Adult 6 Wild 72.3 8.5 8.5 9.8 0.8 0.12 Davis unpubl. data
European toad (Bufo vulgaris) Anuran Adult * Unknown 73.0 18.3 1.3 6.7 0.0 0.25 Friedsohn 1910, cited in Jordan 1938
Fire-bellied Toad (Bombina bombina) Anuran Adult 31 Wild 51.6 25.0 3.9 7.6 12.6 0.48 Wojtaszek & Adamowicz 2003
Rhinella fernandezae Anuran Adult 23 Wild 41.0 19.0 5.0 32.0 1.0 0.46 Cabagna Zenklusen et al. 2011  
African Clawed Frog (Xenopus laevis) Anuran Adult 10 Captive 30.1 26.5 1.2 40.5 1.6 0.88 Hadji-Azimi et al. 1987
Eastern Narrowmouth Toad Anuran Adult 1 Wild 72.3 11.7 0.7 11.7 3.6 0.16 Davis, unpubl. data
Balloon frog (Glyphogloossus molossus) Anuran Adult 18 Wild 41.6 26.3 1.1 8.3 22.7 0.63 Ponsen et al. 2008
Fijian ground frog (Platymantis vitiana) Anuran Adult 40 Wild 66.7 25.3 5.0 15.3 5.0 0.38 Narayan and Hero 2011 Values from animals at time of capture used
Leptodactylus chaquensis Anuran Adult 51 Wild 67.8 25.1 1.5 4.8 0.6 0.37 Attademo et al. 2011 Average values of 3 sites examined
Average for All anurans 63.5 18.7 6.0 9.1 3.3 0.33 Profiles with N/L over 1.0 not included - see below
Standard Deviation 13.1 7.3 4.4 9.3 5.1 0.18 Averages and standard deviations change as new data are added
Eastern Hellbender (Cryptobranchus a. alleganiensis) Urodele Adult 9 Wild 66.5 20.7 4.0 0.0 9.3 0.31 Jerrett & Mays 1973 population from 'Nianuga'
Eastern Hellbender (Cryptobranchus a. alleganiensis) Urodele Adult 18 Wild 60.7 25.9 4.9 0.0 8.4 0.43 Jerrett & Mays 1973 population from 'North Fork'
Eastern Hellbender (Cryptobranchus a. alleganiensis) Urodele Adult 22 Wild 37.5 43.5 14.0 4.2 0.1 1.16 Solis et al. 2007 population from 'MFWR' site , males and females averaged (blood was shipped before smears made)
Eastern Hellbender (Cryptobranchus a. alleganiensis) Urodele Adult 11 Wild 35.5 42.5 18.5 3.2 0.2 1.21 Solis et al. 2007 population from 'EPR' site , males and females averaged (blood was shipped before smears made)
Eastern Hellbender (Cryptobranchus a. alleganiensis) Urodele Adult 31 Wild 49.5 33.0 11.5 5.1 0.8 0.66 Solis et al. 2007 population from 'DR' site , males and females averaged (blood was shipped before smears made)
Eastern Hellbender (Cryptobranchus a. alleganiensis) Urodele Adult 14 Wild 48.0 36.5 3.3 11.4 0.9 0.76 Solis et al. 2007 population from 'CC' site , males and females averaged (blood was shipped before smears made)
Eastern Hellbender (Cryptobranchus a. alleganiensis) Urodele Adult 38 Wild 57.4 28.2 3.7 6.8 1.1 0.49 Huang et al. 2010 Average of 4 populations sampled
Ozark Hellbender (Cryptobranchus alleganiensis bishopi) Urodele Adult 42 Wild 47.2 35.2 12.8 4.2 0.7 0.75 Huang et al. 2010 Average of 2 populations sampled
Red-spotted Newt (Notophthalmus viridescens) Urodele Adult 120 Captive 63.5 24.3 6.2 3.2 2.8 0.38 Bennett & Daigle 1983
Red-spotted Newt (Notophthalmus viridescens) Urodele Adult 13 Captive 55.6 37.6 1.8 0.8 4.2 0.68 Bennett & Johnson 1973
Red-spotted Newt (Notophthalmus viridescens) Urodele Adult 70 Wild 68.2 23.5 0.9 2.2 5.2 0.34 Davis & Erickson, unpubl. data population from one pond in northeast GA
Rough-skinned Newt (Taricha granulosa) Urodele Adult 13 Captive 88.0 7.0 2.0 1.0 1.0 0.08 Friedmann 1970
Triton cristatus Urodele Adult * Unknown 42.9 52.2 3.7 1.2 0.0 1.22 Friedsohn 1910, cited in Jordan 1938
Japanese Newt (Cynops pyrrhogaster) Urodele Adult 23 Captive 3.0 28.0 4.0 57.0 6.0 9.33 Pfeiffer et al. 1990
Red-backed salamander (Plethodon cinereus) - red phase individuals Urodele Adult 35 Wild 68.3 18.0 3.5 9.6 0.7 0.31 Davis and Milanovich 2010 non-breeding, from PA
Red-backed salamander (Plethodon cinereus) Urodele Adult 25 Wild 59.6 35.9 1.5 2.1 1.0 0.88 Davis unpubl. data breeding, from NY
Southern 2-lined Salamander (Eurycea cirrigera) Urodele Larvae 5 Wild 70.9 19.3 1.0 2.9 5.9 0.27 Davis and Milanovich 2010 captured in one stream at Whitehall Forest near Athens GA
Blue-ridge 2-lined Salamander (Eurycea wilderae) Urodele Adult 11 Wild 68.7 22.3 1.5 5.0 2.4 0.32 Davis and Milanovich 2010 captured in one stream at Coweeta, NC
Average for All urodeles

62.1

26.5 4.2 3.8 3.2 0.46 Profiles with N/L over 1.0 not included - see below
Standard Deviation 10.8 8.6 3.0 3.7 3.4 0.20 Averages and standard deviations change as new data are added
Spotted Salamander (Ambystoma maculatum) Ambystoma Adult 11 Reared 31.7 18.1 25.5 24.2 0.6 0.57 Davis and Maerz 2009 Recently metamorphosed, reared at low density
Spotted Salamander (Ambystoma maculatum) Ambystoma Adult 1 Captive 51.1 19.6 19.6 9.8 0.0 0.38 Davis, unpubl. data one individual used in outreach programs
Spotted Salamander (Ambystoma maculatum) Ambystoma Larvae 4 Reared 51.7 14.1 26.9 6.8 0.6 0.27 Davis, unpubl. data approx. 3 mo. old
Tiger Salamander (Ambystoma tigrinum) Ambystoma Adult 1 Captive 46.5 14.0 23.3 16.3 0.0 0.30 Davis, unpubl. data one individual on display in an aquarium
Axolotl (Ambystoma mexicanum) Ambystoma Adult 7 Captive 20.1 21.7 52.0 4.9 1.0 1.08 Ussing & Rosenkilde 1995
Axolotl (Ambystoma mexicanum) Ambystoma Adult 15 Captive 59.0 13.5 22.5 4.0 1.0 0.23 Deparis & Beetschen 1967
Mole Salamander (Ambystoma talpoideum) Ambystoma Adult 34 Wild 41.5 12.7 45.7 0.0 0.2 0.31 Davis & Maerz 2008a
Mole Salamander (Ambystoma talpoideum) Ambystoma Adult 16 Wild 39.0 5.7 51.2 3.9 0.1 0.15 Davis & Maerz 2008b
Mole Salamander (Ambystoma talpoideum) Ambystoma Larvae 45 Wild 52.1 16.7 22.0 8.5 0.7 0.32 Davis, unpubl. data population from one pond in northeast GA
Mole Salamander (Ambystoma talpoideum) Ambystoma Adult 11 Wild 40.5 21.3 32.0 4.9 1.3 0.53 Davis and Maerz 2010 Values from salamanders immediately after capture (reference group)
Marbled Salamander (Ambystoma opacum) Ambystoma Larvae 45 Wild 61.0 13.6 8.5 15.8 1.0 0.22 Davis, unpubl. data population from one pond in northeast GA
Average for All Ambystoma** 47.4 14.9 27.7 9.4 0.5 0.33 Profiles with N/L over 1.0 not included - see below
Standard Deviation 9.3 4.4 12.5 7.3 0.5 0.13 Averages and standard deviations change as new data are added

* Sample size not reported

** Data from Ambystoma salamanders reported separately because of their unusually abundant eosinophils (Davis and Durso 2009)

 

Reference Range for Amphibian N/L Ratios

Amphibian researchers are often faced with the problem of trying to determine if the animals they sampled are stressed, and most of the time there is no prior information on the species under study. However, inferences can be made from the table above, in that a 'universal reference range' can be made for the N/L ratios of all amphibians (below), based on the published reports of all species.

To the left is a histogram of the N/L ratios reported above. There is a clear normal distribution centered around 0.3, with some values appearing outside of the curve, with N/L ratios above 1.0. In the studies representing these outliers, the amphibians appeared to be stressed (i.e. the animals sampled were breeding, or had been transported prior to sample collection, see notes associated with records above).

Then, for any normal distribution of hematological data, it is standard procedure to calculate the reference range as the mean plus or minus 2 standard deviations (Chapter 3, Perspectives in Data Interpretation, in 'Veterinary Hematology and Clinical Chemistry', by Thrall et al. 2006). The mean of all amphibian N/L ratios under 1.0 is 0.34 and the standard deviation is 0.167.

Thus, the reference range of amphibian N/L ratios, in other words the range within which 95% of the population should fall is 0.01 to 0.67.

 

 

Reference List for Amphibian Leukocyte Profiles

Attademo, A. M., M. Cabagna-Zenklusen, R. C. Lajmanovich, P. M. Peltzer, C. Junges, and A. Basso. 2011. B-esterase activities and blood cell morphology in the frog Leptodactylus chaquensis (Amphibia: Leptodactylidae) on rice agroecosystems from Santa Fe Province (Argentina). Ecotoxicology 20: 274-282.

Bennett, M.F., and J.K. Alspaugh. 1964. Some changes in the blood of frogs following administration of hydrocortisone. The Virginia Journal of Science 15: 76-79.

Bennett, M.F., and A.O. Johnson. 1973. Osmotic stress, ACTH and the white blood cell picture in newts, Notophthalmus viridescens. Journal of Comparative Physiology A 82: 333-338.

Bennett, M.F., and K.R. Daigle. 1983. Temperature, stress and the distribution of leukocytes in red-spotted newts, Notophthalmus viridescens. Journal of Comparative Physiology A 153: 81-83.

Cabagna, M.C., R.C. Lajmanovich, G. Stringhini, J.C. Sanchez-Hernandez, and P.M. Peltzer. 2005. Hematological parameters of health status in the common toad Bufo arenarum in agroecosystems of Santa Fe Province, Argintina. Applied Herpetology 2: 373-380.

Cabagna Zenklusen, M. C., R. C. Lajmanovich, A. M. Attademo, P. M. Peltzer, C. M. Junges, G. F. Biancucci, and A. Basso. 2011. Hematology and blood cell cytochemistry of Rhinella fernandezae (Amphibia: Anura) from Espinal and Delta-Islands of Parana River, Argentina. Revista de Biologia Tropical 59: 17-28.

Cannon, M.S., and A.M. Cannon. 1979. The blood leukocytes of Bufo alvarius: a light, phase-contrast, and histochemical study. Canadian Journal of Zoology 57: 314-322.

Cathers, T., G.A. Lewbart, M. Correa, and J.B. Stevens. 1997. Serum chemistry and hematology values for anesthetized American bullfrogs (Rana catesbeiana). Journal of Zoo and Wildlife Medicine 28: 171-174.

Chiesa, M.E., C.E. Rosenberg, N.E. Fink, and A. Salibian. 2006. Serum protein profile and blood cell counts in adult toads Bufo arenarum (Amphibia : Anura : Bufonidae): Effects of sublethal lead acetate. Archives of Environmental Contamination and Toxicology 50: 384-391.

Coppo, J.A., N.B. Mussart, S.A. Fioranelli, and P.A. Zeinsteger. 2005. Blood and urine physiological values in captive bullfrog, Rana catesbeiana (Anura: Ranidae). Analecta Veterinaria 25: 15-17.

Davis, A.K. 2009. Metamorphosis-related changes in leukocyte profiles of larval bullfrogs (Rana catesbeiana). Comparative Clinical Pathology 18(2): 181-186.

Davis, A.K., and A.M. Durso. 2009. Leukocyte differentials of northern cricket frogs (Acris c. crepitans) with a compilation of published values from other amphibians. Herpetologica 65(3): 260-267.

Davis, A.K., and J.C. Maerz. 2008a. Sex-related differences in hematological stress indices of breeding, paedomorphic mole salamanders. Journal of Herpetology 42: 197-201.

Davis, A.K., and J.C. Maerz. 2008b. Comparison of hematological stress indicators in recently captured and captive paedomorphic mole salamanders, Ambystoma talpoideum. Copeia 2008: 613-617.

Davis, A.K., and J.C. Maerz. 2009. Effects of larval density on hematological stress indices in salamanders. Journal of Experimental Zoology 311A: 697-704.

Davis, A. K. & Maerz, J. C. 2010. Effects of exogenous corticosterone on circulating leukocytes of a salamander (Ambystoma talpoideum) with unusually abundant eosinophils. International Journal of Zoology 2010. DOI:10.1155/2010/735937.

Davis, A.K. and J.R. Milanovich. 2010. Lead-phase and red-stripe color morphs of red-backed salamanders (Plethodon cinereus) differ in hematological stress indices: a consequence of differential predation pressure? Current Zoology 56(2): 238-243.

Deparis, P., and J.C. Beetschen. 1967. Comparative observations on some hematological characteristics of diploid and triploid salamanders (Ambystoma mexicanum). Comptes rendus hebdomadaires des séances de l'Académie des sciences. Série D: Sciences naturelles 265: 382-385.

Forbes, M.R., D.L. McRuer, and D. Shutler. 2006. White blood cell profiles of breeding American toads (Bufo americanus) relative to sex and body size. Comparative Clinical Pathology 15: 155-159.

Friedmann, G.B. 1970. Differential white blood cell counts for the urodele Taricha granulosa. Canadian Journal of Zoology 48: 271-274.

Hadji-Azimi, I., V. Coosemans, and C. Canicatti. 1987. Atlas of adult Xenopus laevis laevis hematology. Developmental and Comparative Immunology 11: 807-874.

Huang, C. C., Xu, Y., Briggler, J. T., McKee, M., Nam, P. & Huang, Y. W. 2010 Heavy metals, hematology, plasma chemistry, and parasites in adult hellbenders (Cryptobranchus alleganiensis). Environmental Toxicology and Chemistry 29, 1132-1137.

Jerrett, D.P., and C.E. Mays. 1973. Comparative hematology of the hellbender, Cryptobranchus alleganiensis in Missouri. Copeia 1973: 331-337.

Jordan, H.E. 1938. Comparative hematology, pp. 699-839. In H. Downey [ed.], Handbook of hematology. Vol 2. Harper and Brothers, New York.

Kaplan, H.M. 1952. Variations in white blood cells between normal and red leg frogs. Illinois Academy of Science Transactions 45: 170-176.

Narayan, E., and J. M. Hero. 2011. Urinary corticosterone responses and haematological stress indicators in the endangered Fijian ground frog (Platymantis vitiana) during transportation and captivity. Australian Journal of Zoology 59: 79-85.

Maniero, G.D., and C. Carey. 1997. Changes in selected aspects of immune function in the leopard frog, Rana pipiens, associated with exposure to cold. Journal of Comparative Physiology B 167: 256-263.

Pfeiffer, C.J., H. Pyle, and M. Asashima. 1990. Blood cell morphology and counts in the Japanese newt (Cynops pyrrhogaster). Journal of Zoo and Wildlife Medicine 21: 56-64.

Ponsen, S., N.-a. Narkkong, S. Pamok, K. Sappaso, and W. Aengwanich. 2008. Hematological values and morphological observation of blood cells in balloon frog, Glyphogloossus molossus. Journal of Microscopy Society of Thailand 22: 71-75.

Romanova, E.B., and O.Y. Romanova. 2003. Peculiarities of leukocytic formula of peripheral blood of green frogs under conditions of anthropogenic load. Journal of Evolutionary Biochemistry and Physiology 39: 480-484.

Rocha, G. C., C. M. Ferreira, P. C. Teixeira, D. C. Dias, F. M. Franca, A. M. Antonucci, A. S. Marcantonio, and M. Lauretto. 2010. Physiological response of American bullfrog tadpoles to stressor conditions of capture and hypoxia. Pesquisa Veterinária Brasileira 30: 891-896.

Rouf, M.A. 1969. Hematology of the leopard frog, Rana pipiens. Copeia 1969: 682-687.

Shutler, D., and D. J. Marcogliese. 2011. Leukocyte profiles of northern leopard frogs, Lithobates pipiens, exposed to pesticides and hematozoa in agricultural wetlands. Copeia: 301-307.

Shutler, D., T.G. Smith, and S.R. Robinson. 2009. Relationships between leukocytes and hepatozon spp. in green frogs, Rana clamitans. Journal of Wildlife Diseases 45: 67-72.

Solis, M.E., J.M. Bandeff, and Y.W. Huang. 2007. Hematology and serum chemistry of Ozark and eastern hellbenders (Cryptobranchus alleganiensis). Herpetologica 63: 285-292.

Ussing, A.P., and P. Rosenkilde. 1995. Effect of induced metamorphosis on the immune system of the Axolotl, Ambystoma mexicanum. General and Comparative Endocrinology 97: 308-319.

Wojtaszek, J., and A. Adamowicz. 2003. Hematology of the fire-bellied toad, Bombina bombina L. Comparative Clinical Pathology 12: 129-134.