Mojave rattlesnakes (Crotalus scutulatus scutulatus) lacking the acidic subunit DNA sequence lack Mojave toxin in their venom.

The venom composition of Mojave rattlesnakes (Crotalus scutulatus scutulatus) differs in that some individuals have Mojave toxin and others do not. In order to understand the genetic basis for this difference, genomic DNA samples from Mojave rattlesnakes collected in Arizona, New Mexico, and Texas were analyzed for the presence of DNA sequences that relate to the acidic (Mta) and basic (Mtb) subunits of this toxin. DNA samples were subjected to PCR to amplify nucleotide sequences from second to fourth exons of the acidic and basic subunits. These nucleotide sequences were cloned and sequenced. The nucleotide sequences generated aligned exactly to previously published nucleotide sequences of Mojave toxin. All DNA samples analyzed generated product using the basic subunit primers, and aligned identically to the Mtb nucleotide sequence. However, only 11 out of the 14 samples generated a product with the acidic subunit primers. These 11 sequences aligned identically to the Mta nucleotide sequence. The venom from the three snakes whose DNA did not amplify with the acidic subunit primers were not recognized by antibodies to Mojave toxin. This suggests that snakes with venom lacking Mojave toxin also lack the productive nucleotide sequence for the acidic subunit in their DNA.

Differences in fibrinolysis and complement inactivation by venom from different northern blacktailed rattlesnakes (Crotalus molossus molossus).

Venom from 72 different Crotalus molossus molossus rattlesnakes was examined for fibrinolysis and for their ability to inactivate human complement. The fibrinolytic activity of the venoms was variable, but smaller (younger) snakes had less fibrinolytic activity than larger (older) snakes. Major differences between the venoms was detected by isoelectric focusing, and reflected in the number and pI of the proteins with fibrinolytic activity. Of the 72 venoms tested, ten had no effect and three had low activity on complement. The rest of the venoms strongly inactivated complement. The snakes with no activity on complement measured 55 cm or less in length, except for one snake which measured 53 cm and completely inactivated complement. Two larger snakes (76 and 84 cm) had a reduced complement-inactivating activity. Some venoms strongly hydrolyzed C2, whereas others had mild or no effect on this complement component. The attack on C3 was variable: some had no effect on C3, while other venoms produced a 125,000 mol. wt protein, which was recognized by antibodies to C3. Only mild hydrolysis of C4 was evident in serum treated with some venoms. No relationship was evident between the venom properties of this species and geographical distribution. Venom variability is an important clinical reality, and is an important consideration when attempting to isolate proteases from this snake species for further study.

Detection of alkaline phosphatase in venom by blotting methods.

Venom alkaline phosphatase was detected using a blotting method following electrophoresis. The enzyme gave strong reactions in some venoms, but was absent in other venoms, some within the same species. The mol. wt of the enzyme is close to 100,000 and its pI is between 3.6 and 4.8. The enzyme was inactivated by EDTA and 2-mercaptoethanol, and lost activity by freezing and thawing. Endogenous venom alkaline phosphatase can interfere with alkaline phosphatase-based detection methods. Pre-screening for endogenous venom alkaline phosphatase is recommended prior to using alkaline phosphatase-based detection methods when studying snake venom.

Hemorrhagic and Mojave toxins in the venoms of the offspring of two Mojave rattlesnakes (Crotalus scutulatus scutulatus).

1. The venoms of two Mojave rattlesnakes and those of their offsprings were analyzed for Mojave toxin and hemorrhagic toxin. 2. The venom of one female, collected in Pima County, Arizona, and the venoms of her six offspring contained hemorrhagic toxin but not Mojave toxin (venom B). 3. The venom of the second female, captured in El Paso County, Texas, contained both toxins (A+B venom). Of her 10 offspring, five contained venom with both toxins, two had hemorrhagic toxin only, and three contained neither toxin. 4. Venoms that caused hemorrhage also inactivated complement. A pool of the venoms of the venom B offspring was less toxic than adult pooled venom A.