Biochemical reference intervals for captive bred Crocodylus moreletii and Crocodylus acutus in the Alcuzahue Lagoon, Colima, Mexico

Prado, R.O.F.; Carrillo, D.M.I.; Hernández, R.J.A.; Ruíz, R.J.A.; García, C.A.C.

doi:10.1590/1678-4162-12692

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Veterinary Medicine and Technology and Inspection of Animal Products • Arq. Bras. Med. Vet. Zootec. 74 (06) • Nov-Dec 2022 • https://doi.org/10.1590/1678-4162-12692 linkcopy

Biochemical reference intervals for captive bred Crocodylus moreletii and Crocodylus acutus in the Alcuzahue Lagoon, Colima, Mexico

[Intervalos de referência bioquímica para Crocodylus moreletii e Crocodylus acutus criados em cativeiro na Lagoa Alcuzahue, Colima México]

Authorship SCIMAGO INSTITUTIONS RANKINGS

ABSTRACT

Crocodylus moreletii and Crocodylus acutus are two endangered crocodile species endemic to Mexico. In this descriptive cross-sectional study, a total of 58 crocodiles (30 females and 28 males) were examined to determine and evaluate 24 blood biochemical indicators associated with energy, lipid, protein, mineral and enzymatic metabolic processes. Most of the serum biochemistry parameters were similar among sexes by species. However, male crocodiles showed higher triacylglycerol concentration and alkaline phosphatase activity, and lower globulin concentration than females. There were some significant differences between species. Total bilirubin, triacylglycerol, carbon dioxide, and hydrogen carbonate were higher in Crocodylus moreletii, and urea nitrogen, creatinine, alanine aminotransferase, and aspartate aminotransferase were higher in Crocodylus acutus. These reference values are very important for the protection of crocodiles. The calculated confidence intervals could be used to detect alert situations when at least 5% of the sampled crocodiles would fall outside of the calculated reference interval for a given parameter.

Keywords: blood chemistry; metabolic profile; Crocodylus moreletii; Crocodylus acutus

Analyte*	Unit	Method	Reagent
Energy-lipid profile
Glucose (GLU)	mM	Trinder. GOD-POD¹	1001190^a
Total cholesterol (COL-T)	mM	CHOD-POD. Liquid²	41020^a
Total bilirubin (TB)	μM	DMSO. Colorimetric³	1001044^a
Triacylglycerol (TAG)	mM	GPO-POD. Liquid⁴	41032^a
β-hydroxybutyrate (β-HBA)	mM	Enzymatic⁵	RB1007^b
Protein profile
Blood urea nitrogen (BUN)	mM	Urease-GLDH. Kinetic UV⁶	1001333^a
Albumin (ALB)	g/L	Bromcresol green. Colorimetric	1001020^a
Globulin (GLOB)	g/L	(PROT-T) - (ALB)	Difference
Creatinine (Cre)	μM	Jaffé. Colorimetric-kinetic⁷	1001111^a
Total protein (PROT-T)	g/L	Biuret. Colorimetric⁸	1001291^a
Mineral-electrolyte profile
Calcium ion (Ca²⁺)	mM	Arsenazo III. Colorimetric	CA2391^b
Inorganic phosphate (P_i )	mM	Phosphomolybdate. UV⁹	1001155^a
Sodium ion (Na⁺)	mM	Enzymatic. Colorimetric¹⁰	1001385^a
Potassium ion (K⁺)	mM	UV Test¹¹	1001395^a
Magnesium ion (Mg²⁺)	mM	Xylidyl Blue. Colorimetric¹²	1001286^a
Chloride ion (Cl^-)	mM	Thiocyanate-Hg. Colorimetric¹³	1001360^a
Carbon dioxide (CO₂)	mM	Enzymatic¹⁴	CD127^b
Hydrogen carbonate (HCO₃ ^-)	mM	Enzymatic by CO₂ total and gas dissolved	99852^c
Anion gap (A-gap)	mM	[(Na⁺ + K⁺) - (Cl^- + HCO₃ ^-)]	Difference
Enzyme profile
Alanine aminotransferase (ALT)	U/L	NADH. Kinetic UV. IFCC rec. Liquid¹⁵	41274^a
Alkaline phosphatase (ALP)	U/L	p-Nitrophenylphosphate. kinetic. Liquid¹⁶	41233^a
Aspartate aminotransferase (AST)	U/L	NADH. Kinetic UV. IFCC rec. Liquid¹⁷	41264^a
Creatine kinase (CK)	U/L	NAC. Kinetic UV. Liquid¹⁸	41250^a
γ-glutamyl transferase (γ-GT)	U/L	Carboxy substrate. Kinetic. Liquid¹⁹	41288^a

	Crocodylus moreletii		Crocodylus acutus
	x ± SD	CI	x ± SD	CI	Reference
Energy-lipid profile
Glucose (mM)	3.90±0.47	3.72-4.08	4.02±0.45	3.85-4.19	3.24±0.30^¥
Total cholesterol (mM)	6.14±0.77	5.84-6.43	5.95±0.71	5.68-6.22	6.21±0.94^†
Total bilirubin (μM)	1.24±0.08	1.20-1.27	1.19±0.06	1.16-1.21	1.20±0.47^†
Triacylglycerol (mM)	8.92±0.40	8.76-9.07	7.59±0.35	7.45-7.72	9.02±1.07^†
β-hydroxybutyrate (mM)	0.85±0.14	0.79-0.91	0.79±0.19	0.72-0.86
Protein profile
Blood urea nitrogen (mM)	0.47±0.03	0.45-0.48	0.52±0.01	0.52-0.53	0.44±0.07^†
Albumin (g/L)	26.25±0.32	26.12-26.37	26.31±0.29	26.20-26.43	26.47±3.75^†
Globulin (g/L)	46.11±3.00	44.97-47.25	45.04±3.16	43.83-46.24	44.41±5.85^†
Creatinine (μM)	38.03±1.66	37.39-38.66	39.19±1.66	38.55-39.82	31.19±14.54^†
Total protein (g/L)	72.36±3.22	71.13-73.59	71.44±3.26	70.20-72.68	73.77±7.39^†
Mineral-electrolyte profile
Calcium ion (mM)	3.24±0.18	3.17-3.31	3.26±0.15	3.21-3.32	3.26±0.13^¥
Inorganic phosphate (P_i )	1.70±0.27	1.60-1.81	1.62±0.20	1.55-1.70	1.61±0.20^¥
Sodium ion (mM)	140.19±2.05	139.93-140.46	140.64±3.68	139.24-142.04	141.07±1.95^¥
Potassium ion (mM)	4.37±0.53	4.16-4.57	4.34±0.62	4.10-4.57	4.33±0.40^†
Magnesium ion (mM)	1.11±0.19	1.04-1.19	1.12±0.19	1.05-1.20
Chloride ion (mM)	85.29±4.50	83.57-87.00	84.45±4.47	82.74-86.15	82.74±7.79^†
Carbon dioxide (mM)	26.78±1.02	26.39-27.17	25.85±1.01	25.47-26.24
Hydrogen carbonate (mM)	24.10±0.73	23.82-24.38	23.26±0.74	22.98-23.55
Anion gap (mM)	35.16±6.91	32.53-37.79	37.26±7.80	34.30-40.23
Enzyme profile
Alanine aminotransferase (U/L)	87.81±1.99	87.05-88.57	88.70±1.24	88.22-89.17	89.07±3.72^†
Alkaline phosphatase (U/L)	48.15±4.20	46.55-49.75	48.45±3.38	47.16-49.73	42.10±2.03^†
Aspartate aminotransferase (U/L)	194.09±2.58	193.11-195.08	196.84±1.53	196.26-197.43	193.32±4.64^†
Creatine kinase (U/L)	147.36±8.15	144.26-150.47	148.05±8.08	144.97-151.13	149.22±5.07^†
γ-glutamyl transferase (U/L)	11.00±3.15	9.80-12.20	10.00±3.15	8.80-11.20

	Crocodylus moreletii		Crocodylus acutus
	Male	Female	Male	Female
	x ± SD
Energy-lipid profile
Glucose (mM)	3.93±0.52	3.86±0.43	3.92±0.34	4.10±0.51
Total cholesterol (mM)	6.30±0.68	5.97±0.85	5.74±0.71	6.12±0.70
Total bilirubin (μM)	1.23±0.09	1.25±0.09	1.18±0.07	1.19±0.05
Triacylglycerol (mM)*	9.19±0.19^a	8.62±0.36^b	7.88±0.07^a	7.35±0.32^b
β-hydroxybutyrate (mM)	0.86±0.14	0.84±0.16	0.81±0.21	0.77±0.18
Protein profile
Blood urea nitrogen (mM)	0.47±0.03	0.46±0.03	0.52±0.01	0.53±0.01
Albumin (g/L)	26.29±0.32	26.20±0.32	26.35±0.24	26.35±0.34
Globulin (g/L)*	43.73±3.31^b	48.33±2.66^a	42.64±1.87^b	47.99±1.36^a
Creatinine (μM)	37.69±1.55	38.40±1.76	39.61±1.60	38.84±1.69
Total protein (g/L)	72.96±2.82	71.72±31.72	70.19±3.06	72.46±3.14
Mineral-electrolyte profile
Calcium ion (mM)	3.28±0.16	3.20±0.20	3.21±0.13	3.31±0.15
Inorganic phosphate (P_i )	1.74±0.25	1.66±0.29	1.57±0.20	1.67±0.19
Sodium ion (mM)	140.41±1.74	139.95±2.38	139.31±2.86	141.72±4.00
Potassium ion (mM)	4.44±0.44	4.29±0.62	4.17±0.57	4.48±0.57
Magnesium ion (mM)	1.14±0.13	1.08±0.25	1.06±0.17	1.18±0.19
Chloride ion (mM)	84.51±3.94	86.12±5.06	85.37±4.65	83.70±4.33
Carbon dioxide (mM)	26.76±1.29	26.79±0.66	25.88±1.22	25.83±0.83
Hydrogen carbonate (mM)	24.09±0.92	24.12±0.50	23.30±0.89	23.24±0.63
Anion gap (mM)	36.25±5.89	34.00±7.92	34.81±7.00	39.26±8.00
Enzyme profile
Alanine aminotransferase (U/L)	87.49±1.80	88.15±2.19	88.91±1.51	88.52±0.99
Alkaline phosphatase (U/L)*	51.42±4.47^a	44.63±4.00^b	51.14±1.53^a	46.25±2.83^b
Aspartate aminotransferase (U/L)	194.20±3.31	193.98±1.59	196.66±1.83	196.99±1.29
Creatine kinase (U/L)	146.06±6.42	148.76±9.74	146.33±9.71	149.45±6.48
γ-glutamyl transferase (U/L)	11.59±3.04	10.36±3.24	10.30±2.82	9.76±3.47

	Crocodylus moreletii	Crocodylus acutus
	x ± SD
Energy-lipid profile
Glucose (mM)	3.90±0.47	4.02±0.45
Total cholesterol (mM)	6.14±0.77	5.95±0.7
Total bilirubin (μM)*	1.24±0.08^a	1.19±0.06^b
Triacylglycerol (mM)*	8.92±0.40^a	7.59±0.35^b
β-hydroxybutyrate (mM)	0.85±0.14	0.79±0.19
Protein profile
Blood urea nitrogen (mM)*	0.47±0.03^b	0.52±0.01^a
Albumin (g/L)	26.25±0.32	26.31±0.29
Globulin (g/L)	46.11±3.00	45.04±3.16
Creatinine (μM)*	38.03±1.66^b	39.19±1.66^a
Total protein (g/L)	72.36±3.22	71.44±3.26
Mineral-electrolyte profile
Calcium ion (mM)	3.24±0.18	3.26±0.15
Inorganic phosphate (P_i )	1.70±0.27	1.62±0.20
Sodium ion (mM)	140.19±2.05	140.64±3.68
Potassium ion (mM)	4.37±0.53	4.34±0.62
Magnesium ion (mM)	1.11±0.19	1.12±0.19
Chloride ion (mM)	85.29±4.50	84.45±4.47
Carbon dioxide (mM)*	26.78±1.02^a	25.85±1.01^b
Hydrogen carbonate (mM)*	24.10±0.73^a	23.26±0.74^b
Anion gap (mM)	35.16±6.91	37.26±7.80
Enzyme profile
Alanine aminotransferase (U/L)*	87.81±1.99^b	88.70±1.24^a
Alkaline phosphatase (U/L)	48.15±4.20	48.45±3.38
Aspartate aminotransferase (U/L)*	194.09±2.58^b	196.84±1.53^a
Creatine kinase (U/L)	147.36±8.15	148.05±8.08
γ-glutamyl transferase (U/L)	11.00±3.15	10.00±3.15

	BUN^b	GLOB^c	Cre^d	Ca^2+e	Pi ^f	Na^+g	K^+h	Mg²⁺ⁱ	Cl^-j	A-gap^k	ALT^l	ALP^m	ASTⁿ	CK^o
BT^a	-0.05	0.05	0.09	-0.03	0.05	-0.04	0.03	0.01	0.09	0.04	0.26	0.25	0.26	0.71*
	BUN	0.22	0.46*	0.29	0.24	0.24	0.22	0.23	0.25	0.25	-0.01	-0.02	-0.01	0.20
		GLOB	0.26	0.17	0.20	0.22	0.25	0.22	-0.15	0.27	-0.02	0.67*	0.10	0.26
			Cre	-0.20	-0.24	0.25	0.20	0.23	0.23	0.27	-0.01	0.08	0.17	-0.03
				Ca²⁺	0.81*	0.59*	0.80*	0.61*	-0.50*	0.79*	-0.03	-0.01	0.01	-0.01
					Pi	0.40*	0.87*	0.68*	-0.56*	0.89*	0.01	0.04	0.15	-0.01
						Na⁺	0.24	0.15	0.16	0.64*	-0.03	-0.02	-0.01	0.14
							K⁺	0.65*	-0.54*	0.82*	-0.01	0.01	0.06	-0.02
								Mg²⁺	-0.41*	0.51*	-0.11	-0.06	-0.11	-0.02
									Cl^-	-0.75*	0.01	0.03	0.10	-0.02
										A-gap	-0.01	0.03	0.07	-0.01
											ALT	0.25	0.67*	0.48*
												ALP	0.10	0.10
													AST	0.45*
														CK

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[TFN1] *Official abbreviation of the International Union of Pure and Applied Chemistry; ¹Glucose oxidase-peroxidase; ²Cholesterol oxidase-peroxidase; ³Solubilization with dimethylsulfoxide; ⁴Glycerol-3-oxidasa-peroxidase; ⁵β-hydroxybutyrate dehydrogenase; ⁶Urease-glutamate dehydrogenase; ⁷Sodium picrate; ⁸Copper salts in an alkaline medium; ⁹Ammonium molybdate-phosphomolybdate; ¹⁰Galactosidase-o-nitrophenyl-d-galactopyranose; ¹¹Pyruvate kinase-phosphoenolpyruvate; ¹²Magon sulfonate in alkaline solution; ¹³Mercuric thiocyanate-red ferric thiocyanate; ¹⁴Phosphoenolpyruvate-phosphoenolpyruvate carboxylase-malate dehydrogenase; ¹⁵α-ketoglutarate-glutamate-piruvate-lactate-NADH; ¹⁶Pnitrophenyl phosphate-p-nitrophenol-phosphate; ¹⁷Aspartate-α-ketoglutarate-glutamate-oxalacetate-malate-NADH; ¹⁸Creatine phosphate-ADP-creatine-ATP-D-glucose-ADP-glucose-6-phosphate-NADP-6-phosphogluconate-NADPH; ¹⁹γ-L-glutamyl-3-carboxy-4-nitroanilide-glycylglycine-γ-L-glutamyl-glycylglycine-2-nitro-5-aminobenzoic acid; ^aSpinreact., Girona, Spain; ^bRandox Laboratories., Northern Ireland, United Kingdom; ^cBiolabo Laboratory., Grandcamp-Maisy, France.